Streptomyces Griseus (Krainsky) Waksman and Henrici: A Taxonomic...

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REFtCNCE DO NOT LOAN

STREPTOMYCES GRISEUS

(KRAINSKY) WAKSMAN AND HENRlel

A Taxonomic Study f Some Strains

Technical Bulletin No 1360

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Agricultural Research Service United States Department of Agriculture

To clarify the nature and relationships of the important apt~ioticshyproducing Actinomycetes taxonomic studies were made on a group of 53 streptomycete strains exhibiting the following basic characteristics Spore chains that are straight to flexuous sporulating aerial mycelium colored in tints and shades of yellowish gray (Olive-Buff Ridgway) inability to darken peptone-iron agar or to form brown or black diffusible pigments and ability to utilize o-xylose but notL-arabinosi or L-rhamnose in a chemically defined agar medium Strains studied include some with the epithet labels brasiliensis ctgriseus slreplomycini and uishynaceus and some that reputedly produce keratinase vitamin B12 or one or more of the following antibiotics a~tinomycin complex cycloheximide rhodomycetiu streptocin and streptomytin The basic characteristics of each strain were verified and the following additional characteristics determined optimal temperature range proteolytic activity by six different methods diastatic activity by two methods abilities to reduce nitrate spore-wall ornamentation by electron micrography abilities to utilize six additiOlial carbon compounds sensitivity to lysozyme hy two methods abilities to decompose L-tyrosine xanthine and hypoxanthine and antibiotic activities The results suggest that these strains comprise several subspecies of Streptomyces griseus (Krainsky) Vaksman and Henrici

This taxonomic study (classification and naming) was made as part of the investigations being conducted at the Northern Regional Research Laboratory (NRRL) Peoria Ill on industrial utilization of cereal grains The Xorthern Laboratory is headquarters for the Northern Utilization Research and Development Division The ARS Culture Collection maintained there is one of the worlds largest and most complete collecshytions of industrially important bacteria molds actinomycetes and yeasts The Collection serves as a source of authentic micro-organisms for the fermentative production of organic acids vitamins antibiotics enzymes feeds beverages and foods

Acknowledgment is made to H D Tresuer and L C Davies licroshybiology Department Biochemical Research Section Lederle Laboratories Pearl River XY for supplying Illost of the electron microscope data and to donors of some of the strains used Bristol Laboratories Syracuse NY for cultures of streptomycin-resistant bacteria and Upjohn Co Kalamazoo )Iich for a sample of rhodomycetin This work was supported in part by a grant from the Subcommittee OIl Taxonomy of the Actinoll1ycetes of the Committee on Taxonomy of the American Society for )Iicrobiology

Trade namcs arc used in this publication solely for the purpose of providing specific information )fentioll of a commercial product or company does not constitute a guarantee or warranty of the product by the US Department of Agriculture or an endorsement by the Departshy111ent over other products not mentioned

li

_________________________________________________ _

CONTENTS Introduction_________________________________________________________ _ Materials ad M~t~ods---------- _~_____________bull ___________________ __

P~oteolytlC a~tVlty

Dlastatlc actlvlty ___________________________________________________ _ Reduction of nitrates _________________________ bull _______ _____________ _ Spore morphology and nature of spore surface _________________________ _ Morphology of sporophores and spore chains ___________________________ _ Color _____________________________________________________________ _ Melaninlike chromogenicity _________________________________________ _ Darkening of peptone-iron agar _____________________________________ _ Utiliation of carbon sources for growth _______________________________ _ Sensitivity to lysozyme _____________________________ _______________ _ Decomposition of L-tyrosine _________________________________________ _ Decomposition of xanthine __________________________________________ _ Decomposition of hypoxanthine _________ ___ _________________________ _ Production of antibiotic factors ______________________________________ _

Results _____________________________________________________________ _ General characteristics ______________________________________________ _ Optimal temperature range __________________________________________ _ Proteolytic activity _________________________________________________ _ Diastatic activity __________________________________________________ _ Nitrate reduction __________________________________________________ _ Nature of spore surface ____________________________________________ --Morphology of spore chains ____ bull ____________________________________ _ Colors of aerial and vegetative mycelium ______________________________ _ Melaninlike chromogenicity _________________________________________ _ Darkening of peptone-iron agar______________________________________ _ Utilization of carbon compounds _____________________________________ _ Sensitivity to lysozyme _____________________________________________ _ Tyrosine decomposltion _____________________________________________ _ Xanthine decomposition ____________________________________________ _ Hypoxanthine decomposition ________________________________________ _

DAnti~iotic activity__ bull___ bull ________________________ bull ________ - - _ - - - _ - - --IscusslOn___________________________________________________________ _

Literature cited ______________________________________________________ _

fge 1 1 6 7 7 7 7 7 8 8 8 8 8 8 9 9

10 10 10 11 11 11 11 13 13 14 15 15 16 16 17 17 19 25 29

Washington DC_ Issued November 1966

iii



A T cxonomic Study cf Some Strains

By A J LYONS JR and 1 G PRIDHAM

Northern ltilization R~search and Development Division

Agdcultural Research Service

iNTRODUCTION A large number of streptomycetes in the ARS Culture Collection have

been divided into groups based on morphology color of aerial mycelium melaninlike chromogenicity (ability to form brown deep brown or black diffusible pigments) and ability to utilize D-xylose L-arabinose and L-rhamnose Particular groups have the additional characteristic that they contain strains which reputedly produce different antibiotics One group was selected for further taxonomic study in an effort to characshyterize the strains more precisely and to determine the relationship between the different antibiotic-producing strains The group selected contains strains that have flexuous spore chains that form pale-yellow to grayish-yellow [ISCC-NBS Xos 89 and 90 (9) 1 Olive-Buff (22)] aerial mycelium that are nonchromogenic and that have the ability to utilize D-xylose but not L-arabinose or L-rhamnose

All the strains of Slreplomycc8 griseus (Krainsky) Waksman and Henrici (2 10) in the ARS Collection that have been reported to produce streptomycin fell into the group selected The group also includes a few strains of S griseus reported to produce cycloheximide rhodomycetin and streptocin and strains with the specific epithets brasiliensis streplomycini and rvinaceus as well as some strains isolated at the Northern Division

This work further demonstrates the close relationships of these strains Classical taxonomic procedures allow the differentiation of these strains and the 1915 culture of Streplomyces yriseus Waksman and Henrici The strains studied were separated into several categories (subspecies)

MATERIALS AND METHODS The taxonomic procedures used with the strains studied (as listed

below) are either cited or described The stock cultures and inocula for the various strains were prepared as outlined by Lyons and Pridham (14) The designations specific epithets and histories of the streptomycete strains selected from the ARS Culture Collection at the Northern Regional Research Laboratory (iIERL) are given in table 1

1 Italic numbers in parentheses refer to Literature Cited p 29

1

2 STREPTOMYCES GiUSEUS A TAXONOMIC STUDY

TABLE l-Des-ignabions specific epithets and histories of selected streptomycete strains from Ihe AR S Culture Collection

Designation Antibiotic(s) or used Specific epithet other metabclic

activity

BaldaccL_ __ __ __ brasiliensis_____ - (1)

ACTU 60L__ ___ griseus_ _ _ ______ streptomycin____ _

B-128L________ grisells_________ (1)

IMRU 3475_____ grisells _________ kemtinase _______ _

IPV 423x_______ griseu8_________ (I)

NCIB 8225 _____ griseuL________ (1)

NCIB 900L ____ griseus _________ streptomycin phage resistant

History of strains received by NRRL

Received 1S54 from E Baldacci Instituto Patologia Vegetale Universita di Milano Italy (IPV) Received by Baldacci from Instituto Superiore di Santa Rome Italy as an isolate from human disease

Received 1955 from H Sakai ~~kaguchi Laboratories Department of Agrishycultural Chemistry Faculty of Agriculture Tokyo University (ACTU) as No 601

Isolated 1950 at NRRL from Japanese soil

Received 1961 from R Gordon Institute of Applied Microbiology Rutgers University (lMRU) as 3475 from US Patent 2988488 (23) 61361

Received 1959 from E Baldacci IPV as strain 17 for International Common Experiment (12) ex IPV 423x ex ATCC (American Type Culture Collection Strain No unknown)

Received 1958 from National Collection of Industrial Bacteria (NCIB) as NCIB 8225 British Drug House Ltd 1950

Received 1958 from NCIB as NCIB 9001 Pamela M Boyd MRC Antishybiotics Research Station Clevedon (No R25) 1952 Actinophage resistant and streptomycin producer

See footnote at end of table

3 TECHNICAL BULLETIN 1360 US DEPT AGRICULTURE

TABLE I-Designations specific epithets and his~vres of selected streptomycete strains from the ARS Culture Collection-Continued

Designation Antibiotic(s) or History of strains used Specific epithet other metabolic received by NRRL

activity

NCIB 9004 _____

S-1471- ________

SAW 4-L ______

SAW 4-2 ______

SAW 4-3 _______

SAW 4-4 _______

SAW 4-5 _______

SAW 4-6 _______

SAW 4-7 _______

SAW 9-L ______

griseus _________

griseuB- ________

griseus _________

griseus _________

griseus _________

griseuB- _______

griseus _________

griseu8 _________

griseus _________

griseus _________

B12 ___ -- _ --- ---- shy

(I)

streptomycin cycloheximide

streptomycin streptocin

streptomycin______

streptomycin______

streptomycin______

streptomycin______

(1)

streptomycin______

Received 1958 from NCIB as NCIB 9004 M Lumb Boots Pure Drug Co Ltd (No FD 196) 1952 Production of vitamin B12bull

Isolated 1950 at NRRL from Liberian soil

Received 1957 from ATCC as ATCC 10137 ATCC received it from S A Waksman (SAW) as strain 4 derived from strain 3463 the original streptomycin-producing stram with orIginal strain No 18--16

Received 1961 from R Gordon as SAW 3496 same as Vaksman 4 a colony isolate of SAW 3463 (18-16)

Received 1958 frorr Torry Research Station Aberdeen as NCIB 8506 Waksman 4 ATCC 10137

Received 1950 from J C Sylvester Abbott Laboratories as Abbott 2 2k-13 derived from S griseus Waksman 4

Received 1950 from J C Sylvestedas Abbott 3 sl-18 as erived from S griseus Waksman 4

Received 1950 from J C Sylvester as Abbott 5 m-365 as derived from S griseus Waksman 4

Received 1958 from NCIB as NCIB 8232 (mutant from Waksmans strain 4) M Lumb Boots Pure Drug Co Ltd (No FD 40) 1951

Received 1957 from ATCC as ATCC 11429 as SAW 9 (3463) Rutgers University 1953

Bee footnote at end of table

4 STREPTOMYCES GRISEUS A TAXONOMIC 1lJigtY

T ABLE I-Designations specific epithets and histories of selected streptomycete stroins from the ARS C1tlture Collection-Continued


activity

SAW 9-2 _______ griSelt8 _________ streptomycin Received 1951 from D cycloheximide Gottlieb as Gottlieb

A-2 from Waksman 9 SAW 9-L _____ griselts _________ (1) Received 1959 from NCIB

as NCIB 8237 (mutant from Waksmans strain 9) M Lumb Boots Pure Drug Co Ltd (No FD 124) 1951

griselts _________SAW 3464 ______ streptomycin______ Received 11)50 from S A Waksman a8 SAW 3464 D-1 isolated in 1944 from the throat of a chicken as orginal streptomycin producer

SAW 3481 _____ grisclts _________ streptomycin ______ Received 1950 from S A Waksman as SAW 3481 a freshly isolated streptomycin-produling culture

SAW 3495 ______ _____griseus rhodomycetin Received 1950 from S A pink variant Waksman as SAW

3495 a natural variant of S grileus

griseus ________SL 842 _________ _ streptomycin______ Received 1954 from F Carvajal Schenley Laboratories as SL 842 calpable of producllg 200 to 500 micrograms of streetomycin per milliliterSL 2060 ________ griselts _________ (1) Received 1946 from G W Ward Schenley Labozdtories as SL 2060

NIlIJ SM-L ___ griseus var anti-Gram-positive Received 1962 from Y rhodochrous antibiotic Okami National

produced (not Institute of Health streptomycin) Tokyo Japan (NIHJ)

as SM-lNIHJ SM-2 ____ griseu8 var anti-Gram-positive Received 1962 from Y

rhodochrous antibiotic (not Okami NIHJ as SM-2 streptomycin)

NIHJ SN-J-L __ griseus var streptomycin______ Received 1962 from Y rhodochrou8 Okami NIHJ as

SN-I-J NIHJ SN-J-2___ griseus var streptomycin ______ Received 1955 from T

rhodochrous Yamaguchi University of Tokyo Japan who received It from Y Okami NIHJ as SNmiddot-1-J


________________

________________


TABLE I-Designations specific epithets and histories of selected streptom1jcete strains from the ARS C1llture Collection-Continued

Designation used

NIHT SN-2 (2) __

NIHJ SN-14-L_

NIHJ SN-14-2__

Lilly L _________

Lilly 3 __________ Lilly 5 __________ Lilly 7 __________ SAW 3479 ______

ARI1780_______

NI9003 ________

Carpenter_______

E______________

Specific epithet

griseus var rhoigtiJhroU8

grise us var rhodochrous

griseus var rhodochrou8

sp _____________

sp _____________ sp _____________ sp _____________ sp _____________

streptomycini____

vinaceus________

Antibiotic(s) or other metabolic

activity

streptomycin______

streptomycin______

streptomycin______

streptomycin_____ _

streptomycin_____ _ streptomycin_____ _ streptomycin_____ _ actinomycin

complex

(1)

(1)

(1)

(1)


Received 11)62 from Y Okami NIHJ as SN-2 (2)

Received 1962 from Y Okami NIHJ as SN-14

Received 1955 from K Saito Institute of Fermentation (IFO) Osaka Japan as IFO 3358 who received it from Y Okami NIHJ as SN-14

Received 1950 from J M McGuire Eli Lilly amp Co as Lilly boil isolate not necessarily S griseus but believed to produce streptomycin

Do Do Do

Received 1955 from S A Waksman as SAW 3479 isolated in the Waksman Laboratory in 1948 and belonging to the S coelicolor group (35) Also lis ted as 34-1 (antagonist) (personal communication to T G Pridham 2662)

Received 1962 from V D Kuznetzov Antibiotics Research Institute (ARI) USSR Moscow as strain 1780 a Krasilnikov culture

Received 1953 from Nagoa Institute (NI) Japan as NI 9003 Actinomyces S-20 of K Saito

Received 1959 from C C Carpenter Syracuse University Research Foundation labeled as coiled verticilate

Received 1960 from K Crook Bristol Laboratories Syracuse NY as strain E

See footIgtote at end of table

----------------

----------------

6 STREPTOMYCES GRISEUS A TAXONOMIC STUDY

TABLE I-Designations specific epithets and histories of selected streptomycete strains from the ARS Cult1lre Collection-Continued

Designation Antibiotic(s) or History of strains used 8pecific epithet other metabolic received by NRRL

activity

8-62___________ (1) Isolated 1953 at NRRL--------------shyfrom 8ap 8alvador soil sample 8-90L _________ (1) Isolated 1953 at NRRL from Illinois soil sample

8-1757 _________ (1) Isolated about 1957 at NRRL as B-6-6

8-1758_________ (1) Isolated about 1957 at NRRL as B-27-1

--------------8-175L ________ (1) Isolated about 1957 at---------------

NRRL from Australian soil sample

8-1760_________ (1) Isolated about 1957 at NRRL from 80uth African soil sample

---------------

8-176L ________ (1) Isolated about 1957 at NRRL as F-3

-------------shy8-1762 _________ (1) Isolated 1958 at NRRL

from California soil sample

--------------shy

8-1763 _________ (1) Isolated 1959 at NRRL from Alaska soil samte

--------------shy8-1764 _________ (1) Isolated 1959 at NRR

from Illinois soil sample

---------------shy

8-1765 _________ ________________ (1) Do8-1766 _________________________ (I) Do8-1767 _________________________ (1) Do

1 To date there has been nothing reported about antibiotic activity or other metabolic activity that might or might not be present

Proteolytic Activity

The proteolytic activity of each strain was determined by six methods (1) cultivation in 15 percent plain gelatin (Difco) in tubes in which the inoculum (02 ml of a 48-hour tryptone-yeast extract (TYE) broth culture) was placed on the surface of the substratum No stab was made Cultures were incubated at 28deg to 30deg C for 14 days On the 14th day each culture was refrigerated at 3deg to 5deg for 1 pour and then examined for liquefaction and color of diffusible pigment (2) cultivation ~n 15 percent plain gelatin and 1 percent soluble starch (Difco) with inoculation incubation and reading as in the first method (3) the Waksman 15 percent plain gelatin-dish method with incubation at 18deg to 20deg (32) (4) the Waksman 15 percent plain gelatin and 1 percent starch-dish method with incubation at 18deg to 20deg (32) (5) the Gordon and Smith casein-dish method with incubation at 28deg to 30deg (6) and (6) the Gordon and Mihm gelatin-hydrolysis method with incubation at 28deg to 30deg (4) Suitable uninoculated control media were used for comparisons

7 TECHNICAL BULLETIN 13(0 US DEPT AGIUCULTUHE

All cultures were observed at 11 days and the degree of liquefaction or hydrolysis of gelatin degree of clearing of caseill agar and nature of diffusible pigments were recorded

Diastotic Activity

Two methods were used to detect diastatic activity of each stmin (1) the method outlined in Lyons and Pridhal11 (14) and (2) the Gordon and lihnl starch-hydrolysis method modified by use of soluble starch (Difco) and substitution of Lugols iodine solution for 05-percent ethanol (1)

Reduction of Nitrates

The methods are based on procedur(s outlined by the Subcommittee 011 thl Taxonomy of the Actinomycetes (26) and by the Committee on Bacteriological Technic (3) Cultures were tested and obseled for reduction of nitrate after 14 days growth at 28deg to 30deg C

Spore Morphology and Nature of Spore Surface

Electron 1l1icrogrnphs of most of the strains were obtained from Tr(sner and others Electron micrographs of the rest of the straills were oLtained at the Xorth(rn Division also by the procedure of Tresner and others (29) Formvar-coated grids were impressed on the aNial mycelia of 14-day inorganic salts-starch (ST) agar dish cultures

Morphology of Sporophores and Spore Chains

~rorphological studies WCIf made as outlined ill reports by Hesseltine and others (7) Lyons and Pridham (14) and Pridluul1 and others (19) except that nutrient agar was not used

Color

fethods for determination of color$ of sporulating aerial mycelium reverses of cultures and difTusible pigments are cited alld explained ill Lyons and Pridham (14) In addition to thfse obsClvations were noted with potato slants and slices Potato slants were pleplued and inoculated as outlined previously (14) Also peeled white potatoes were cut into slices and soaked in distilled water at 3deg to 5deg C for 24 hours The slices were drained placed in petri dishes and sterilized for 1) to 20 minutes at 121deg Artel sterilization a small amount of sterilized distilled

~ water was added to each dish to prevent the potato slice from drying out Each slant and lice w(re inoculated as outlined (14) After 14 days incuhn tion at 28deg to 30deg each preparation was examined and colors of aerial my(eliulH vegetative growth and potato were recorded

Colors wer~ initially keyed out according to Ridgway (22) Later the Ridgway col( l tabs were matched to their nearest equivalent in the Color Harmlly )Innual 4th edition (27) Colors of aerial mycelia were keyed OUf to the appropriate color series in the system of Pridham ancl others (9 and in the one proposed by Tresner (28) Finally all trivial names of color tabs selected were converted to their lSCC-NDS equivalents (9)

8 STREPT0IYCES GHISEUS - TAXONOilIC STUDY

Melaninlike Chromogenicity

The ability of each strain to produce b1O1I deep brown 01 black diffusible pigments was determilled by observation of TYE broth culshytures gelatin cultures and potato cultures after appropriate incubatioll periods

Darkening of Peptone-Iron Agar

The method for this determination is cited and outlined by Lyons and Pridham (14)

Utilization of Carbon Sources for Growth

The method for determining carbon utilization patterns of each of the strains is also outlined by Lyons and Pridham (14) Carbon sources used were n-xylose I-arabinose L-rhamnose D-glucose D-fructose D-galactose raffinose D-mannitol i-inositol and salicin (all Difco prodshyucts) Ability to utili~e sucrose was determined by cultivating each strain on Czapeks solution (CZ) agar for 14 days at 28deg to 30deg C

Sensitivity to Lysozyme

Two methods were used to determine the sensitivity of the strains to lysozyme (1) the method described by Gordon and Mihm (5) and (2) a method based on that of Smolelis and Hartsell (25) III the second method 10 m1 of TYE broth was inoculated with 2 loopfuls of spores from 14-day-old tomato paste-oatmeal (TPO) agar or yeast extract (YE) agar slant cultures and incubated for 48 hours at 28deg to 30deg C on a rotary shaker Each culture was centrifuged the supernatant was discarded and the cells were adjusted to give a reading of about 10 percent light transmission with a Lumetron colorimcter (red filter) by addition of glycerol broth (5) Lysozyme (Nutritional Biochemicals Co Cleveland Ohio) solution was prepared according to directions given in Gordon and Mihm (5) except that 10 m1 was added to 90 m1 of glycerol broth Five milliliters of this mixture was added to 5 ml of the adjusted cell suspension and changes in light transmission were noted at 20-minute intervals for 2 hours The lysozyme-cell suspensions were incubated at 28deg to 30deg during the tests A few additional trials were made at incushybation temperatures of 37deg and 45deg Cells of Micrococcus lysodeikl1cUS Fleming NRRL B-287 were used as a control

Decomposition of L-Tyrosine

The method outlined by Gordon and Smith (6) was used to determine whether any of the strains could decompose L-tyrosine Cultures were observed after 14 days incubation at 28deg to 30deg C for clearing of the substrate as an indication of decomposition

Decomposition of Xanthine

The method outlined by Gordon and Mihm (4) was used to determine whether any of the strains could decompose xanthine Cultures were

9 TECHNICAL nUlLETIN 1360 US DEPT AGRICULTURE

observed after 14 days incubation at 28deg to 30deg C for clearing of the substrate as an indication of decomposition

Decomposition of Hypoxanthine

The method outlined by Gordon and lIihm (5) was used to determine whether any of the strains could decompose hypoxanthine Cultures were observed after 14 days incubation at 28deg to 30deg C for clearing of the substrate as an indication of decomposition

Production of A~tibiotic Factors

The antibiotic-producing capacity of each strain was determined by the Waksman agar streak method by cross-antagonism tests and by paper-disk assays and paper-strip chromatography of shaken-flask and static fermentation filtrates and mycelial extracts

The llledium used for the Waksman agar streak and cross-antagonism tests was a medium (SCG) based on medium A-4h of Warren and others (88) as outlined in Lyons and Pridham (14) with 15 percent agar added The primary streaks were incubated for 7 days at 28deg to 30deg C and at that time the bacteria yeast molds and streptomycetes were streaked at right angles to the growth The dishes were held an additional 3 to 5 days and the zones of inhibition were recorded Test strains used for agar streak determinations were Bacillus subtilis Cohn emend PrasJlIowski XRRL B-765 Escherichia coli ()[igula) Castellani and Chalmers XRRL B-766 E COI1 XRRL B-27-18 (streptomycin resistant) E coli XRRL B-2422 (streptomycin resistant) E coli NRRL B-1079 (streptomycin dependent) Staphylococcus aUleus Rosenbach NRRL B-313 S au reus NRRL B-2747 (streptomycin resistant) Mycobacterium sp XRRL B-692 Candida albicans Robin (Berkh) NRRL Y-477 11ucol lamann-ianlls Ioel XRRL 1839 and a strain of S alllellS NRRL B-313 whose resistance against rhodomycetin was developed for use in this study

Shaken-flask fermentations were run in five media for each strain These media were (1) the A-4h (SeG) broth of Warren and others (38) for 4 days (2) the Pridham and Gottlieb chemically defined basal broth (18) containing 1 percent D-glucose and 01 percent yeast extract (Difco) for 4 days (3) the cycloheximide production medium No 26 of Whiffen (39) for 4 days (4) the rhodolllycetin production medium of Shockman and Waksman (24) for 6 days and (5) the streptomycin production meshydium No 25 of Whiffen (39) for 6 days The rhodomycetin production medium also was used in a static fermcntation in order to detect strepshytocin production This medium was arbitrarily selected because the medium used for the production of streptocin was not clearly defined in the two papers by Waksman and associates (36) and Kupferberg and coworkers (11)

Shaken-flask media were inoculated with 5 percent (vv) of 48-hour TYE broth cultures of the streptomycetes Seeded flasks were incubated on a Gump rotary shaker (except those for streptocin production which were held under static conditions) operation at 200 rpm at 28deg to 30deg C On the fourth and sixth days the mycelium was separated from the liquor by filtration and centrifugation The mycelium was washed with distilled water and separated into two portions idethanol (10 m1

10 HlHEPlO~(YCE mUSEUS A (AXO-O~C HTUDY

of an 80-percent methanol solution) was added to one portion and the mixture was ground in a tissue grinder and filtered thlOlIgh Whatman Xo 1 filter paper This material comprised the methanol extmct Ethyl ethel (10 m of anhydrous ether) was added to the second pOl-tion and the mixture was shaken for 30 seconds The ether was decanted Five milliliters of distilled water US added to the lllytJelium and the mixture was ground in a tissue grinder Two more ethel extractions (10 m each with aO-second shaking) were made of the mycelium and the three ethel extracts were combined After the ether was allowed to evaporate the residue was taken up in 10 1111 of 95-percent ethanol to provide the ether-ethanol extracts

The culture filtmtes methanol extmcts and ether-ethanol extracts were assayed against six test organisms by the paper-disk assay method The test organisms for the paper-disk assays were B subtilis XRRL B-765 B coli fRRL 13-766 Mycobacterium sp XRRL B-692 Sarcina lutea RRL 13-1018 C albicans XRRL 1-middot477 and ill mlnannian1ls XRRL 1839 These organisms were grown in I-7 agar (20) except for ill mmannian1ls which was glOwn in Iucor synthetic agar (VISA) a chemically defined medium (20)

Paper-strip chromatography studies were carried out on all filtrates methanol extmcts and ether-ethanol extracts with three solvent systems These were water-saturated butanol butanol-saturatpd water and 10 percent NH4Cl All paper strips were spotted with 150jLl of sample and sealed in a glass chromatography jar After equilibmtion for 1 hour solvents were added to the jars and the strips developed by des~ending chromatography The strips were removed from the jars when the solvent fronts approached 1 inch from the bottom of the strips The fronts were marked and the strips were inverted and allowed to air-dry for 1 hour At that time the strips were laid on seeded agar trays for 1 hour in order to permit diffusion of the active factors into the agar Test organisms used were B sublilis KRRL B-765 and Saccharomyces pastOlianllS Hansen NRRL Y -139 Trays were then incubated at 28deg to 30deg C until growth was sufficient to locate zones of antibiotic activity Control strips were used in all jars

RESULTS

General Characteristics

All the strains studied exhibited the general characteristics of the genus Streptomyces All grew weU aerobically were of the same size (mycelium about O5jL to 10jL in diameter) and formed spores in chains with more than three Rpores per chain

Optimal Temperature Range

All the strains were able to grow well at temperatures of 18deg 25deg and 28deg C Ai 37deg growth was limited and a few strains were unable to grow NOlle of the strains grew at 45deg or 55deg Strain Baldacci which was received with the amppecific epithet brasiliensis and reportedly isolated from human diseas) required the same temperatures for growth as the rest of the strains t hat were isolated from soil



All the strains studied were proteolytic when tested by the six methods used Based on these and other experiments the methods most suitable were the Waksman 15 percent gelatin-dish method and the Gordon and Smith casein-dish method The Waksman 15 percent gelatin-dish gives well-defined zones of liquefaction and there is no need to refrigerate cultures before they are read With the cultures studied the zone diameters ranged from 18 to 41 mm The average zone diameter was 33 mm The Gordon and Smith casein-dish method gives rapid weIlshydefined clearing and the medium is simple to prepare Also the caseinshydish method far excels methods based on whole milk and litmus milk as liquid culture media for determination of action on casein Zone widths with the casein-dish method were 23 mm with some of the strains Some cultures completely cleared the medium The average width of cleared zones was 30 mm

Diastatic Activity

All the strains were diastatic The ST agar cultures when flooded with Lugols iodine solution gave no indication that any starch was left in the agar according to the method used (14)

Results were difficult to interpret when the Gordon and Mihm starch-agar cultures were flooded with 95-percent ethanol according to directions With Difco soluble starch and 95-percent ethanol no detectshyable zone limits could be seen Control dishes of the starch agar became only faintly cloudy when flooded with 95-percent ethanol Duplicate cultures when flooded with Lugols iodine gave readily discernible zone limits and control dishes of the agar became deep blue These results may represent differences in reaction to 95-percent ethanol between the potato starch used by Gordon and Iihm and the Difco soluble starch With Lugols iodine reagent the widths of the decolorized zones on the Gordon and Iihm media cultures ranged from 20 mm or wider Some strains gave activities that represented complete hydrolysis of the starch

Nitrate Reduction

Table 2 shows that neither the chemically defined broth nor the organic-based broth gave uniform results Strains from the same origin (eg SAW 4-1 and SAW 4-2) gave different results In a number of instances the same strain allowed detection of nitrite in one tube of a pair but not in the other (eg SAW 3495 in the organic-based broth) Obshyviously nitrate reduction tests as used with streptomycetes require modification and improvement

Nature of Spore Surface

Electron micrographs of each of the strains listed show that the organisms all have smooth-walled spores The morphology of the spore surfaces was quite uniform The spores are ellipsoidal and measure approximately 05J1XlOJl (fig 1)


TABLe 2-Reduction of nitrate by strains of streptomycetes 1

Designation used

BaldaccL________________________ ACTU 601______________________ B-128L _________________________ IMRU 3475______________________IPV 423x________________________ _ NOIB 822L_____________ ________ NCIB 9001 ______________________ _ NCIB 9004 ______________________ _ S-1471_________________________ SAW 4-L_______________________ SAW 4-2________________________ _ SAW 4-3_________________________ SAW 4-4________________________ _ SAW 4-5 ________________________ _ SAW 4-6_______________________ __ SAW 4-7_________________________ SAW 9-1 ________________________ _ SAW 9-2_________________________ SAW 9-3 ________________________ _ SAW 3464________________________ SAW 3481 ______________________ _ SAW 3495 _______________________ _ SL 842___________________________ SL 2060__________________________ NIHJ SM-1____________________NIHJ SM-2_____________________ _ NIHJ SN-J-1 __________________ _ NIHJ SN-J-2 ___________________ _ NIRJ SN-2 (2) __________________ _ NIHJ SN-14-L _________________ _ NIHJ SN-14-2 __________ ________Lilly I _________________________ _ Lilly 3___________________________Lilly 5 __________________________ _ Lilly 7 __________________________ _ SAW 3479 _______________________ _ ARI 1780_______________________ _NI9003_________________________ _

S-62____________________________ _~~~e~~~~=======================8-901___________________________ _ S-1757__________________________ _ 8-1758__________________________ _8-1759__________________________ _ 8-1760__________________________ _ 8-1761__________________________ _8-1762__________________________ _ 8-1763__________________________ _ 8-1764__________________________ _ 8-1765 __________________________ _ 8-1766__________________________ _8-1767__________________________ _

Chemically defined broth

Tube 1 Tube 2

+ + C+) (+)(+) (+) C+) C+)

(-)

C+) (-)C+) (-)

(+) (+)

(-) (-) (-) (-)

C+) (+)

(+) (+)

(-) (-) (+) (+)

(-)

(-) (-)

(-) (-)

(+) (+)(+) C+)(- ) (- )

C+) (- )(+) (-) (- ) C+) C+)(-) (- )

C+) (+) (- ) (- )

(- )

c+) (+) + +

Organic-based broth

Tube 1 Tube 2

(+) + +

(+) (+) + + - -

(+) (+)c+) (+)

(+) + +

c+) C+)(- ) c+) (+)c-) C-)c+) C+)(- ) c-)(- )

+ +(+) (- ) + + - (+)

(+) + + + + -

(- ) (-) (- )

(- ) (-) (- ) C+) C+)(+) + -C+) C+)

c+) (+) + + + +

C+) c+)c+) c+) + +

(+) c+)

c+) C+)(- ) c-) + + + +

(+) (+)C+) (+) + + + +

(8ee footnote on facing page)

bullbull bull bullbullbull

- -

13 TECHxrCAL neLLET1X 1360 ( DEPT AGRICULTUHE

B

(p S D

FrIHllB 1 ~[tJrph()lflgy of SPOrtS and nature of surfares of some sl rains of SireploshyIIIfIimiddotI bull lriwI~ l ()riinal lJlii (ullure (~IU 126) B SA )-1 (~[RC 1middotIG Ilrigintl st rrptomyrin-proliucing Cuiturc ( SAW l4J5 rhodomycetinshyproliuing (ull un IJ IU l7-O Actill)mlc(~ srlflloIIJJrini Electron microscopy of figures Hand ( by II D irctiner and ~l C Davies ~[agnification about G)OOX to SOOOX

Morphology of Spore Chains

All tlninlitCd WPlC lC]Cgated to section HCetus-F]Cxibilis (Ill) (19) aft(gttmiddot in situ ohslvatic)]ls of pehi-dih cultUlPs At low magnification thp (haill of POlCS appear us tufts with lplativcly little branching The POlP (huins arp pPIlNully flCxtious and ralely straight aii showll in figure 2

Colors of Aerial and Vegetative Mycelium

All trw cuI turps had aerini lllyceliuIll colored 1Il tints and shades of yellow (tahle 3)

(Footnote to table 2)

I ymhols u~ed - nitrates not rfduced (-) slight pink color produced on addition of rpap(ntH 1+1 palp r(middotd (0101 prod1((d on addition of reagents + deep red color produ(((l Oil addit ion of rpuglnts (lilt iVlited on organie-lmsed lind chemically defin((] IIwdia (14 days at 2) to ~() C bull a-dimethylnnphyillmine lillifanilie acid and Zn dust reagents)

TREPIO~IYCES GHISE1S A TAXOXOlIlC STtDY

FIGClIE 2-~licromorphol()IY of a representative strnin (~I lOO3) of Streptomyces

grisels (ulture Iron for H days at 210 to ~Oo C on inorganic salts-starch agar

Tints and shadrs of yrllowish pillk and orange yellow are often associshyated with till charnctrristic yrlloll colors rxhibitcd by the aerial mycelia of thr culturrs gtuclird This characteristic apprars true not only for thosr culturrs whosr rgrtatir Illycrliulll is yellow to yellowish brown but also for thosr culturs IIhosr ngrtativp l11ycrliulll is rrd to purple 1hr yrllowish pink and omng( y(llow arr mol( prollounced with those eulturrs with r(ci to purplr egtatin Illycelium This dual color phenomrnon has bren not(d with (special frequency when strains arc culturNi Oil TPO agar Th( phrnomellon suggests that the cultures can be subdhidrd into two color rat(gorirs has(cl on thr color of their aerial mycelium On til( one hand wlllll the cultures w(re compared side by sid( the color ciifTrr(nces w(r( not sufficirnt to allow objective separation into two categori(s 011 tlH other hand wh(n the cultUl(s were compared on thr hasis of th( color of tlllir vrg(tatin mycelium it as relatively simple to organiz( strains into two eat(gori(s (yrllow to y(llowish brown and red to purplC) as showll in tablr 4


011( of til( gtmillg produced brow11 de(p brown or black diffusible piglllllts with TYE broth glatin tubs gelatin dishrs potato slants or potato Jie( Strain with 1((1 to purp( r(v(lses ho((I produced difTusib( piglllrllts in pink or iolt Beeaus no othel difTusiblc pigmrnts ofiI1lportancr w(r( lIotNl it is cOllclu(Cd that all strains studied arc nOllchromogcnic within the limits of our definition


TABLE 3-Colors of aerial mycelium of strains grown for 14 days at 28deg to 30deg C1

Tresner ISCC- ISCC-Ridgway name and

Ridgway CUM color name and

Tresner name and

color wheel

Pridhllm color

NBS color

NBS color

plate 2 group desigshynation 3

desigshynation 4

series (28)

series (19)

names of Ridgway

tabs 5

names of CUM tabs 5

Olive-Buff O-yy Biscuit Parch- Yellow Olive- Pale Yel- Grayish XL 2ec ment buff lowS) Yellow

1Hdb 90 Pale Olive- O-yy ivory 2db ivory 2db Yellow Olive- Pale Yel- Pale Yel-

Buff XL buff low S9 lowS) Grayish Yellow 90

Deep Olive- O-YY Bamboo None Yellow Olive- Grayish Grayish Buff XL 2gc buff Yellow Yellow

)O 90 Tilleul- O-Y Sand 3cb Bisque Red Red or Pale Yel- (No

Buff XL 3ec olive- lowish name) buff Pink 31

Pale o-y Pearl lba Pearl pink Red Red or Pale Yellowish Pinkish 3ca olive- Orange White Buff buff Yellow 92 XXIX 73

1 The color of the aerial mycelium of eueh of the strains studied is represented by one or more of the color names listed Colors determined from examination of ST agar dish cultures

2 Ridgway color designations representing the closest approximation that could be made

3 Color name and designation from Color Harmony Manual (CHl) (137) representshying the closest approximation that could be made for the Ridgway tabs

4 The closest CHi color name and designation that could be made by using the color tabs su~gested by H D Tresner (2S) at the workshop on streptomycetes held during the Eighth International Congress of llicrobiology at Montreal Canada August IS 1962 (17)

S The Inter-Society Color Council-Xational Bureau of Standards OSCC-NBS) Circular 553 (9)


Xone of the strains exhibited a bluish-black coloration of the subshystratum when gr9wn on peptone-iron agar There has been some question whether this test actually detects hydrogen sulfide for which it was devised (J S) however the results appear to correlate with those for the usual tests for melaninlike chromogenicity

Utilization of Carbon Compounds

All the strains exhibited a similar carbon utilization pattern (table 5) except for their activities on salicin Xo direct cOITelation could be made between other charac~eristics and salicin activities

_____

________

16 STREPT0[YCES GRISEUS A TAXONOlUC STUDY

TABLE -t-Colors of Legetative mycelium of strains grown for 14 days at 28deg to 30deg Cl

Yellow to yellow-brolL7 reverses Strains ATCU 601 B-1281 IMRU 3475 IVP 423x XCIB 8225 NCIB 9001 NCIB

9004 SAW 4-1 SAW 4-2 SAW 4-3 SAW 4-4 SAW 4-5 SAW 4-6 SAW 4-7 SAW 9-1 SAW 9-2 SAW 9-3 SAW 3464 SAW 3481 SL 842 SL 2060 Lilly l Lilly 3 Lillv 5 Lilly 7 ARI 1780 Carpenter E S-62 S-901 S-1757 S-1758S-1760 8-1762 81763 8-1764 S-1765 8-1766 and 8-1767

Ridgway CH~I name and ISCC-NBS ISCC-NBS name and plate 2 designation 3 color names of color names of

Ridgway tabs laquo CHiI tabs

Chamois XXX _ Honey Gold 2ic _ Ioderate Yellow 87 i[oderate Yellow 87 Cinnamon-ButT Lt Amber 3ic Moderate Yellow 87_ Ioderate Orange

XXIX Moderate Orange Yellow 91 Yellow 71 Dark Oran~e

Light Yellowish Yellow 72 Brown 76

Colonial ButT Pastel Yellow Light Yellow 86 ____ Moderate Yellow 87 XXX 1Yzfb

Cream-ButT XXX_ Bamboo 2fb_ ____ iIoderate Yellow 87_ Moderate Yellow 87 Light Yellow 86 ____ Pale Yellow 89

Deep Chrome III Brite Yellow 3na __ Stron~ Orange Stron~ OrangeYellow 68 Yellow 68

Honey Yellow Mustard Gold 2ne_ Moderate Yello87_ Dark Yellow 88 XXX Dark Yellow 88

Isabella Color lIustnrd Gold 2pe_ Dark Grayish Deep Yellow 85 X-XX Yellow 41

Mustard Yellow lIaize 2hb Strong Yellow 84 ___ Moderate Yellow 87 XVI

Naples Yellow Lt Wheat 2ea ____ Light Yellow 86 ____ Light Yellow 86 XVI

Ochraceous-ButT Amber 3Ic ________ Light Yellowish (No name) XV Pink 28

Moderate Yellowish Pink 29

Olive-Brown XL __ Clove Brown 3111_ Grayish Yellowish Dark Yellowish Brown 80 Brown 78

Tawny-O live Topaz 3ne ________ Moderate Yellowish (No name) XXIX Brown 77

Light Olive Brown 94

(See footnotes on facing page)


Ve were unable to obtain reliable data on sensitivity to lysozyme when the Gordon and Iihm (5) method was used The results in table 6 suggest the same difficulties are experienced as with the nitrate-reducshytion tests Uniform results were obtained when a method based on that of Smolelis and Hartsell (25) was applied All strains were sensitive to lysozyme under these test conditions Solected results are presented in table 7 Some strains were lysed rapidly others less so Also duplicate preparations gave similar results

Tyrosine Decomposition

All strains used in this study decomposed L-tyrosine


TABLE 4-Colors of tegeiatiue mycelium of strains grown for 14 days at 28deg to sao Cl-Continued

Red to purple reverses

Strains Baldacci 8-1471 SAW 3495 NIHJ S11-1 NIHJ SM-2 NIHJ SN-J-l NIHJ SN-J-2 NIHJ SN-2(2) NIHJ SN-14-1 NIHJ SN-14-2 SAW 3479 NI 9003 S-1759 8-1761

Ridgway CHM name and ISCC-NBS ISCC-NBS name and plate Z designation 3 color names of color names of

Ridgway tabs 4 CHM tabs

Brownish Vina- Dusty Coral Light Grayish Dark Pink 6 ceo us XXXIX 6Ygc Red 18 Dark Yellowish

Pink 30 Dahlia Carmine Raspberry 9pe ____ Dark Purplish (No name)

XXVI Red 259 Dark Maroon- Raspbery Wine Dark Reddish Dark Purplish

Purple XAI (gtpg Purple 242 Red 259 Very Dark Purplish

Red 260 Dark Vinaceous Cedar 6gtlile_______ Grayish Red 19 ____ Grayish Red 19

XXVII Deep Corinthian Antique Rose 7Ie __ Grayish Red 19 ____ Grayish Red 19

Red XXVII Deep Livid Brown Old Wine 7Yng___ Grayish Red 19 ____ (No name)

XXXIX Indian Lake Raspberry 9pc ____ Moderate Purplish Moderate Purplish

XXVI Red 258 Red 258 Grayish Purplish Dark Purplish

Red 262 Red 259 Purplish Vina- Dusty Rose 7Ygc_ Light Grayish Dark Pink 6

ceous XXXIX Red 18 Vinaceous-Purple Raspberry 9ne ____ Moderate Purplish (No name)

XXXVIII Red 258 Grayish Purplish

Red 262

I The color of the veetative mycelium (reverse of cultures) of each of the strains studied is represented in one of the color names listed Colors determined from examination of Inlernational glycerol-asparagine agar dish cultures

Z Ridgway color designations representing the closest approximation that could be made

3 Color name and designation from Color Harmony Manual (CRM) (amp7) representshying the closest approximation that could be made for the Rideway tabs

4 The Inter-Society Color Council-National Bureau of Standards (ISCC-NBS) Circular 553 (9)

Xanthine Decomposition

All the strains that had ye1low to yellowish-brown vegatative myceli11m decomposed xanthine Differences were noted with the strains that have red to purple vegetative mycelium (table 8)

Hypoxanthine Decomposition

AU the strains decomposed hypoxanthine

18 STREPTOlIYCES GRISEUS A TAXONOMIC STUDY

~rABLg 50~nilizati(n of carbon compounds by strains of streptomycetes 1--_ 0 sectIl Cll Cll Cllen00 rn Cll rn

en Cll 0 0 0Designation used Cll-t rn Cll Il Il pound Cll CIl CIl0 B 0 02 0~ ~ 0 0 E E 0 0 Ill degiil Il

01 gt l Il0 a 01 01 0

~Il 0 f8 Il sect ~ 1 ~ 0 01 ~ OJZ A A ~ I ~ A 70 U1

aldacci __________________B - - I + - -TCU 60L _______________ + + (=) + + -A-1281 ___________________ - + + - + + - + - (+)BMRU 3475_______________ - I + + c-) - + + - + - (+)

I - (- ) -VP 423x _________________ + + + + - + - c+)1 cm 8225 _______________ - + + - -- + + - + - (- )N - c-)lCIB 9001 ______________ + + - + + - + - +- + c-) shycm 900L ______________ + - + + + - c+)N-1471 ___________________ - + + - - + + - + - c-)8 - + + (- ) - + + - + - c+)AW 4-1 ________________8AW 4-2 _________________ - + t - - + + - + - c+)8AW 4-3 _________________ - + - - + + - + - c+)8 + (- ) + c+)AV 4-4 _________________ - + - + - + shy8AW 4-5 _________________ - + + - - + + - + - c+)8 - + + (- ) - + + - + - (- )AV 4-6 _________________8 -- + + c-) - + + - + - (- )AW 4-7 _________________ 0~V9-1

S _________________ - + + - - + + - + - (- )SAW 9-2 _________________ - + + - - + + - + - +S - + + (-) - + + - + - (+)AW 9-3 _________________8AW 3464 ________________ - + + - - + + - + - c+)8 - + + - - + + - - (+)AW 3481 _______________ +S - + (- ) - + - - c+)AV 3495 ________________ + + +S - T + - - + + - + - c+)SL 842 __ - + (-) - - - (+)L 2060 ---~------------ + + +__________________ +S - + + - - + + - + - c+)TTTTJ SM-L ____________N - (- ) (- ) - (- ) - - (+)NUrT SM-2 _______________ + + +

- + + (- ) - + + - + - (+)NIHJ SN-J-L ___________ - (-) - C+) - (+)IHJ SN-J-2 ____________ + + + + -N - + + (- ) - + + - + - (+)NTHrT SN-2(2) ____________ - + (+) (- ) - C+) + - + - (+)NIHJ SN-14-L ___________ - (+) c-) - - shy+ + + + c+)NUTJ SN-14-2 ____________ -illy L __________________ - + c+) (-) - + + - + (+)L 01y 3____________________ - + + - - + + - + - c+)L - c-) -Lilly 5 ____________________ - + +

(- ) + + - + - c+)

Lilly 7 ____________________ + + - + + - + - c+)- + C-) +SA W 3479 ___ 0 __________ + - + - + - C-) - -ART 1780_________________ + + - + + - + - +- + + - - + + - + - c+)NT 9003 __________________

Carpenter _________________ - + + (- ) - + + - + - (+)- + + - - + + - + - (- )

~------------------------_____________________ + - + - + (+)S-62 - + - + shy(- ) shyS-901 ____________________ - + + - + + + - (- ) (-) shy8-1757 ___________________ - + + - + + + - (- ) - Ii8-1758 ___________________ - + + - + + - + - c-)

S-1759 ___________________ - + + (- ) - + + - + - c-) 8-1760 ___________________ - + + - - + + - + - c-) 8-1761 ___________________ - + C+) - - + + - + - c+) 8-1762 ___________________ - + C+) - - (+) + - + - (+) 8-1763 ___________ - _______ - + + - - + + - + - c-) S-1764 ___________________ - + + - + + - + - (+)

- + + - + + - + - c+)C==)18-1765_ - - bull -___________________ - - - - - - __ - - ---OJ + - + + - (+)8-1766 - + + shy8-1767 ___________________ - + + - + + - + - C+)

- + - - (+) + - (+) - (- )I+ I shy

(See footnote on facmg page)

------------------

TECHNICAL BULLETIN 1360 US DEPT AGRICULTURE 19

TARLE G-Sensitiuity to lysozyme of strains of streptomycetes as determined by the Gordon and J[ihm (5) method 1

Growth in Growth lysozyme broth

Designation used in control

Tube 1 Tube 2

Baldacci___________________________________ _ Poor (- ) (+)SAV 4-1 _________ ______________________ bull __ GoodSAV 9-1 __________________________________ _ Good (- ) (+)SAV 3495 ________________________________ _

HJ SN -~-L _____________________________ _ Good + +~I GoodARI 1780_ bull _______________________________ _ + +GoodNI 9003 _______________ ___________________ _ Good + +

t Symbols used - good growth (not sensitive to lysozyme) (-) less growth (+) growth present hltt poor + no growth detected (sensitive to lysozyme)

TABLE 7-Sensitivity to lysozyme of strains of streptomycetes as determined by a method based on that of Smolelis and Hartsell (25) 1

Light transmission throu~h culture after addition of lysozyme at 28deg to 30deg C for-

Designation used 0 20 40 60 80 100 120

min- min- min- min- min- min- minshyutes utes utes utes utes utes utes

Per- Per- Per- Per- Per- Per- Per-

BaldaccL_____________________ SAV 4-1 ______ bull ______________ SAW 9-L ___________________ SAW 3495 ____________________ NIHJ SN-J-L ______ bull ________ ARI1780 _____________________ Nf 9003 ______________________

cent 22 22 18 22 23 19 20

cent 50 72 34 39 36 30 57

cent 66 81 51 61 62 49 64

cent 73 82 66 73 76 67 70

cent 75 83 76 76 75 77 73

cent 76 88 81 80 76 84 77

cent 0 85 82 81 81 87 80

1rIicrococclLS lysoceikticlls NRRL B-287 2

20 ca 95 96 96 96 96 96

t Readings made with a Lumetron colorimeter (red filter) 2 Used as a control

Antibiotic Activi~y

It was possible to identify those strains that produce streptomycin through use of the Vaksmall agar streak method paper-disk assays of cdlture filtrates and cross-antagonism tests Streptomycin-dependent

(Footnote to table 5) t Priclhllm ancl Gottlieb basal agar 10 clays at 28deg to 30deg C Symbols used -

no growth (-) faint growth probably no utilization (+) poor to fair growth + good growth and positive utilization

20 SlREPTOMYCES GRISEUS A TAXONO~IC STUDY

TABLg 8-Decomposition of xanthine by strains of streptomycetes with red to purple vegetative mycelium 1

Clearing of Designation used xanthine

medium 2

-------------------------------------------------1---------BaldaccL__ ____ _ _ __ ___ ___ ___ __ ____ ____ __ __ _ ____ _____ ____ ____ _ _ _ + 8-1471________________________________________________________ + 8AV 3495________________________ -______ -_____________________ + NIHJ 8Zvl-l_ _ ___ ____ ________ __ __ __ ____ __ ___ ____ _ _ _____ ____ __ _ _ + NIHJ 811-2 __________________________________________________ _ NI1IJ SN-J-1______________ - __________________________________ _ NIfTJ SN-J-2_________________________________________________ _ NIHJ SN-2 (2)_______ _______ ___ ___ __ ____ __ __ ___ _ _ _ _ _ ___ _ _ _ __ _ _ _ + NllfJ SN-14-1 ________________________________________________ bull NIHJ SN-14-2________________________________________________ _ S~V 3479_____________________________________________________ 4shy~I 9003_______________________________________________________ 4shy8-1759________________________________________________________ 4shy8-1761____________ bull _____________________________ _ _ _ ___ __ __ ___ +

1 Gordon and Mihm (4) method 14 days at 28deg to 30deg C Symbols used +medium cleared and xanthine decomposed - medium not cleared and xanthine not decomposed

2 Results from duplicate tests

and resistant bacteria and grisein- and rhodomycetin-resistant bacteria were especially helpful as test organisms in this regard In table 9 are presented the results of agar streak tests obtained with strains representshying variOllS subcategories finally established in the work All the known streptomycin-producing strains and 15 other strains allowed the streptoshymycin-dependent strain of Escherichia coli to grow The rest of the strains under ftudy did not The 1915 isolate of Actinomyces griseus IMRU 3326 (34) was included in these tests to determine whether it produced streptomycin although it does not exhibit the basic characteristics of the group of strains under study There are several reports in the literature that also suggest the inability of this strain to produce streptomycin (J 21 37) Only slight antifungal activity was detected with the 1915 bull strain

Cycloheximide was presumptively identified by activity of the strains against a yeast and a mold in the agar streak tests and paper-disk assays Presumptive confirmation of identity was made by paper chromatogshyraphy and knowledge of the histories of the strains

In an attempt to determine whether any of the strains could produce streptocin culture mycelia were extracted with ether A simplified proccdure based on that described by Waksman and others (36) and on the solubility of streptocin in ethyl ether and ethanol was used with each strain Because streptocin is reported to have activity against Gramshypositive rods and cocci acid-fast bacteria yeasts and molds activity of such fractions against these organisms was used to presumptively detect streptocin

Under the test conditions mycelial extracts of the original streptocinshyproducing cu1ture (IMRU 3533) gave activity against four of the five different types of organisms when the culture was grown in a streptoshymycin-production medium but not in the streptocin-production medium


TABLE 9-Antibiotir actiLity of representative strains as determined by the lVaksman agar streak method with SCG medium incubated at 28deg 030deg CI

Designation used

01 01 1

S TisCU8 I 0 0 0 o o 7 S IH5 culture

SAW 4-1 2 14 21 o 20171 01 011 0 8)streptomycin cycloheximide I

ARI liSO bullbullbullbull _ ______ (3) 30 o 20SAW 3495 ___________ 1 3~ ~i ~ olgl 2~ O~ 22 3S oBaldaccL __________ i 01 OJ 0 07 47 0 5 0 o oSAW 4-2 ____________ 32 0 0 01 241 14 28 32 o 20 NI 9003 bullbull ___ - ---- --I o 0 0) 01 01 0 5 15 o o NIHJ S1-2 _________1 o 3t 0 31 3 3 4 7 o o NIHJ 8~-J-L __ __ _ o 0 0 0 0 0 o 0 o o8-1761______________ 10 13 8jl03011 20_-1 6 12125301 o oS-1762 ________ middot _____1 Of 011 0 4 OJ 0 5j 515 o o

I I i1

1 Figures indicate length of inhibition zones in millimeters except for those for the streptomycin-dependent strain

2 Figures indicate millimeters of growth (not inhibition) extending out from the priml1ry streak

3 Complete inhibition of bl1cteril1 under test conditions

or any of the other media used Trace activity suggestive of streptocin also was obtained when the strain was grown in the cycloheximideshyproduction medium The results with those strains whose ether-ethanol extracts showed antibiotic activity are presented in table 10 The original streptocin producer is included as well as two other strains reported to produce this antibiotic Preliminary paper chromatographic study of the mycelial extracts from the streptomycin-production medium suggests that the antifungal activity detected is polyenic in nature and is not cycloheximide as originally reported In view of Yamaguchi and Saburis (40) work showing that many different kinds of streptomycetes exert activity against trichomonads there is some question as to the precise identity of the antitrichomonal factors reportedly produced by strains other than DIRU 3533 So far strain BIRU 3533 has been reported to produce streptocin cycloheximide a second antitrichomonal antishybiotic l and a factor active against Gram-positive cocci Our preliminary

- -

22 STREPIOIYCES URlSEU A fAXONOlIC STUDY

results suggest that the antifungal activity is not cycl)heximide Strain SAV 3490 a knowll streptomycin producer also has been reported to produce streptocin Oil the basis of anti trichomonal activity of a sublimable fraction (11) In our hands only questionable evidence for the production of streptocin based on activity of extracts against Gram-positive rods and cocci was obtained Xo activity was detected against the acid-fast organism used Results of a sinlilar nature were obtained with strain DIRU 340~ reported by toamaguchi and Sabmi (40) to exert antitrichshyomonal activity

The presumptive detectioll of rhodomycetin was based on activity of this antibiotic against Gram-positive rods and cocci acid-fast bacteria and a rhodomycetin-resistant strain of SaphyiococctlS (IWmiddotWS

The antibiotic activity of representatie strains detected in samples from shaken-flask fermentations with SeG medium is shown in table II The results given arc only illustrative In some instances other media gave bettel activity In table 12 results are presented for one strain (SAW 4-1) when cultivated in all of the media studied Fair antifungal activity was noted with many strains in the agar streak test However

TABLE 1O-Strains showing antibiotic actiLity in the ether-ethanol elracls oj Uw mycelium when grown in the slreptodn-prochlciion medium as determined by the paper-disk method 1

00 0middot_to=-tshy-I 70- ~=lDesignation used ~~

N - ~~ - ~~Czc ~z CQ UJ

SAW 4-L ____________ _ SAW 4-22 _____________ _ + + SAW 4-4 _____________ _ SAW 4-7 _____________ _ + + + SAW 9-1 2____ bull ________ _ + SAW 9-3 _____________ _ SAW 3481 ____________ + + NCIB 9004 ___________ _ + + Lilly 7 ________________ _ + + ACTU 601 ___________ _ + E____________________ _ + Carpenter_____________ _ + + S-62 _________________ _ + + S-1761 _______________ _ + + 8-1760 _______________ _ + + 8-1764 _______________ _ + 8-1765 ________________ + + 8-1766______________ _ + NIHJ SN-2(2) ________ _ + IMRU 3533 3__________ _ +

+ antibiotic activity detected - antibiotic activity not dmiddotected 2 Strains reported to produce strcptocin 3 Original streptocin-producing strain

l


this activity did not appear in assays of samples from the shaken-flask and static fermentations X0 single medium ga e the best activity fOI all strains although SeG seemed to be a superior one

Cross-antagonism studies of all the strains gave results as illustrated in table 13 The 191) culture of Streptomyces griseus was included in these studies and found to be sensitive to streptomycin and to rhodoshymycetin These results further suggest that this strain does not belong to the streptomycin-producing group Although the rhodomycetin-proshyclucing strain did not inhibit streptomycin-producing strains it was inhibited by them

Strain ABI 1780 inhibited strain SAW 9-1 however none of the other streptomycin-producing strains inhibited SAW 9-1 X0 explanation as to the cause of this inhibition can be advinced except that strain ARI 1780 was extremely active OIl all agar streak tests

Paper-strip chromatography was run with all of the fermentation samples and extracts flom each of the strains used in this study Represhysentative results are ShOl] in ligures 3 and 4 All the strains gave patshyterns limilar to those illustrated if antibiotic activity were present Standard samples of streptomycin and cycloheximide were run at the same time and compared with the activities shown here From these studies it was concluded that the antibiotic activities produced by many of the strains are streptomycin and cycloheximide Another factor (designated factor X in fig 3) was found to be present in the mycelial

TABLE ll-flntibiolic acti6ty of representaliue streptomycete strains tn shaken-jlask fermentations as determined by paper-disk method 1

ci 00 ~

ltltnCl -lt ~t ~g s~ 0~Cgt ~Cgtor- ~I-- lt0 8~lt-gt1 -I t 1 7 1 sectgz

Strain designation and fermentation products lQ ~Q I ~I Sgtlt ~

~l il ~l ~l lt~ sectl~~ ~~ g~ ~ ~~ ~~ ~ ~~ 6~~~

~~

~Z ltZ ~Z t Z EZI ~Z ~ l1 i tQ c) ~

Baldacci ISCG filtrate_ ___ bull ______ bull ________________ -- ---- 0 14 0 () 0 0 SCC melhan(ll extract - _ _ ______ - __ - - __ - - - - - -_ 0 15 0 14 0 0 SCG ethanol-ether extrnct _______________________ n n 0 0 0 0

SAW 4-1 SeG filtrate_ _______________________ ----- ---- 0 21 32 15 0 0 SeG methanol extracL _________________________ 0 0 0 14 0 0 SeG ethanol-ether extracL ______________________ 0 0 0 0 0 0

)1 HJ S~1-2 SeG filtrate_ _____________________ bull _______ - ---I 15 14 0 00 0SeG methanol extracL _________________________ 0 (2) 0 00 0 SCG ethanol-ether extract ________________ - _---- 0 15 0 (2) 0 0

SAW 3495SeG filtrate _____________________________ - ---- shy 0 0 15 0 0 0 SeG methanol extracL _________________________ 0 0 0 0 0 0 SeG ethanol-ether extracL ______________________ 0 0 0 0 0

I 0

1 Figures indicate diameter of inhibition zones in millimeters 2 Trace

24 STREFTmlYCES GRISEUS A TAXONOMIC STUDY

TABLE 12-Antibiotic activity of strain SAW 4-1 (reported to produce streptomycin and cycloheximide) in shaken-flask and static fermentashytion8 as determined by the paper-disk method 1

ci 00 ~ g 5 g Cj r- ~~Ogt 0 ~r-

or- -r- ~Ogt I -I 1 til sect1 ~ltl

Media ampnd fractions tested Q sectQ r Q ~Q 5gt-i ~e1 ~~ ~ E~~~ t~-~ l~ ~~ ~~ ltl~ g~ ~ ~ ~8~ ~~~~ ~~ gZ cs Z ~Z aZ ~Z 8 ZcslQ Xl CIlt c ~

SCG medium 2Fittrate _______________________________________ Methanol extracL _____________________________ Ethanol-ether extract __________________________

0 0 0

21 0 0

32 0 0

15 1lt 0

0 0 0

0 0 0

Pridham and Gottlieb 2Filtrate_______________ bull _______________________ ~Iethanol extract_______________________________ Ethanol-ether extract _________________ bull ________

0 0 0

18 0 0

26 0 0

(3) 0 0

0 0 0

0 0 0

Cycloheximide production 2Filtrate_______________________________________ ~Iethanol extract_______________________________ Et11anol-ether extracL__________________________

0 0 0

20 0 0

32 0 0

16 14 0

0 0 0

0 0 0

Rhodomycetin production 4Filtrate___ ___________________________________ ~Iethanol extract _______________________________ Ethanol-ether extract ___________________________

0 0 0

22 0 0

35 0 0

19 0 0

0 0 0

0 0

(3) Streptomycin production 4Filtrate_______________________________________ 0 0 0 0 0 0

~Iethanolextraet_______________________________ Ethanol-ether extraeL__________________________

0 0

18 18

0 0

0 0

0 0

0 0

Streptocin production 5Filtrate_______________________________________ 0 20 30 17 0 0 ~ethanolextract_______________________________ Ethanol-ether extract __________________________

0 0

15 14

0 0

(3) (3)

0 0

0 0

1 Figures indicate diameter of inhibition zones in millimeters 2 Incubation 4 days at 28deg to 30deg C on rotary shaker 3 Trace 4 Incubation 6 days at 28deg to 30deg C on rotary shaker 5 Incubation ) days at 28deg to 30deg C in static culture

extracts of many of the streptomycin producers This factor might be streptocin The chromatographic pattern as shown in figure 3 is similar to the pattern obtained with the oily residue isolated from fermentations of strain IlIRU 3533 according to procedures of Waksman and others (36) Because no standard antibiotic sample of streptocin was available we cannot conclusively identify the activity as streptocin

All the strains with red to purp1e vegetative mycelium gave patterns similar to those of the rhodomycetin-producing culture and to the strains labeled S griseus var 1hodochrous with one exception The patterns are recorded in figure 4

Seven strains did not produce streptomycin were sensitive to a streptoshymycin-producing culture and also had yellow to yellow-brown vegetative mycelium Two had antiyeast activity other than cycloheximide as

-- -- ---- ----

25 lECHNICAL BULLETIN 1360 US DEPl AGUlCULTURE

TABLE l3-Results of cross-antagonism study with representative stmins 1

[Waksman agar streak method with 8CG medium at 28deg to 30deg C]

~ I C)l= L) 7 0 Cjl Cgt Z 00Primary streak I t- IfJ IfJ lt3 M

0 0 Cgt N M cgtdesignation used ~ aS 0 lta

11) 0 Cgt 1shy t t Cgt ~ 0 1 IfJ IfJ

lt -lt 7 lt Z Z l Z IfJIfJ IfJ

I 1915 Cultule ______________ () () 0 () () 0 0SAW O-L ________________ 25 () 11 4 to ---shy

0 () l a 6 5SAW 4-2 _________________ 30 () 0 () 8 7 5 6 8 4 3ARI 1780 _________________ 32 10 8 () 20 18 12 13 17 17 118-1762 ___________________ ---- 4 5 ) 0 0 () 0 0 0 2SA W 3495 ________________ 20 () 0 () () () U () 0 () IINIHJ SM-2 ______________ 15 5 () () () I) () 0 0 () 0

NIHJ SN-J-L ____________ 15 () 0 () () () 0 0 () 0 ()Baldacci __________________ 0 () () lJ () 0 () 0 0NI9003 __________________ 0 () () 0 () 0 0 0 () ()8-1761 ___________________ --~-I () I

---- 5 7 7 [) 3 0 0 3 4 0

1 Figures indicate length of inhibition zones in millimeters

shown in figuret One strain produced antibacterial activity that gave chromatographic patterns sO bulllewhat similar to streptomycin The sensitivity of the strain to streptomycin along with its chromatographic pattern (fig 3) suggests that the activity was not due to streptomycin

The four other strains in this group did not produce any antibacterial activity However three of these strains gave activities similar to cycloheximide when filtrates and extracts were chromatographed One strain was inactive

Filtrates and extracts made with the original 191) strain of S gliseus were chromatographed (fig 4) This strain showed no antibacterial activity but gave an antifungal pattern Ultraviolet absorption curves indicated this activity was due to a mixtUle of hexaene and heptaene antifungal agents

We were unable to confirm the reports of Umezawa and others (30 31) and Okami (15) that some of the strains with red to purple reverses produce streptomycin None of the criteria we used suggested this possibility

DISCUSSION Om study confirms in general reports on the taxonomy of Streptomyces

griseus from other laboratories (16 33 37) Further these and related studies now in progress suggest that S gliseus can be so subdivided into taxa by classical taxonomic methods that certain relationships between the taxonomic characteristics and the qualitative nature of the antibiotics produced by given strains become evident Identification of individual strains with individual antibiotics reported within each group however cannot be accomplished by classical procedures with much confidence Carbon utilization patterns have been especially helpful For example


Oritil

front wsa asw 10 wsa asw 10 wsa asw 10

HH4CI HH4CI HH4CIStreptomycin Cycloheximide factor X Test orlanism Test orlanism Test arianism

B subtiis Sacco pastorianus B subtilis

Streptomycin-producinl strains Iyellow to yellow-brown reverses)

Orilin


HH4CI HH4CI HH4CICulture HRRL a-1281 Culture S-1760 Culture S-901 Test arianism Test orlanism Test orlanism

Sacco pastorianus Sacco pasforianus B subtiis

Nonstreptomycin-producinl strains Iyellllw to yellow-brown reverses)

FIGURE a-Chromatographic pattern categories obtained with filtrates and mycelial extracts of strains of Streptomyces griseus Solvent systems WSB watershysaturated butanol BSW butanol-saturated water and 10 percent ammonium chloride


Orilil

Front WSI ISW 10 WSI ISW 10 WSI ISW 10

NHCI NHCI NHCI Rhodomycetin Culture SNmiddot14middot2 Culture Smiddot1161 Test orlanism Test orlanism Test orlanism

8 subtiis 8 subtilis 8 subtiis

Cultures with red to purple reverses

bull

WSI ISW 10 NHCI

Orilinal 1915 Culture SAW 3326

Test DrlanisM M l-amonnionus

FIGURE 4-Chromatographic pattern categories obtained with filtratetl and mycelial extracts of strains of Streptomyces griseus

28 STREPTO~iYCES GRISEUS A TAXONOlIIC STUDY

ali those strains of S gr-iseus in our collection reported to produce strepshytomycin were placed in the collection of strains at present under studyHowever specific information on the total qualitative antibiotic-proshyducing potential of strains is needed to make further separations withconfidenceThe results of the present study based only on criteria in the classicalsense ie excluding criteria concerned with production of antibioticssuggest there are only two distinguishable taxa within the collection ofstrrins examined These are (1) those strains with yellow to yellowishshybrown vegetative mycelia and reverses and (2) those strains with red topurple vegetative mycelium amI reverses These could be designatedas two different subspecies of S griseus In accordance with the Intershynational Code of Komenclature of Bacteria and Viruses (8) the taxonStreptomyces griseus (Krainsky) Waksman and Henrici subspecies griseus(Krainsky) Waksman and Henrici would automatically be createdAs it turns out these three taxa can be neatly differentiated by classicalt ltonomic methods eg colors of vegetative mycelium or reverses andflrbon utilization patterns However the broad taxonomic studiesconducted at the Northern Division indicate that beyond these relativelysimple differentiations collections of strains identified in this manner canbe further subdivided although not with confidence unless one resortsto more highly sophisticated procedures eg either objective determinashytion of the qualitative nature of the antibiotics or possibly the pigmentsproduced by given strains It appears that the best way to handle theproblem of classification and 110menciature of streptomycetes is to utilizethe species-subspecies concept and to take into account that antibioticproduction represents an important objective characteristic of thesubspecies Our work thus far suggests that each of the three possiblesubspecies defined along classical lines does in fact contain strains whichproduce different antibiotics and hence can be further subdivided Atthe present stage of our work the following treatment of the strainsstudied can be made Streptomyces gliseus (Krainsky) Yaksman and Henrici subspeciesgliseUs (Krainsky) Waksman and Henrici strains with flexuous chains of 1spores spores smooth-walled yellow to yellowish-gray aerial myceliumoccasionally tinged with pink yellow to yellowish-brown vegetativemycelium nonchromogenic utilizes D-glucose D-xylose L-arabinoseD-fructose D-galactose raffinose D-manl1itol i-inositol and salacin butnot L-rhamnose when incorporated in the Pridham and Gottlieb chemshyically defined basal agar (J8) produces hexaene and heptaenc antifungalantibiotics and no as yet detected antibacterial antibiotic(s) Proposedneotype strain nIRU 3326 the 1915 isolate of Waksman and Curtis(34) which also is proposed as the neotype strain of Actinomyces griseus 4Krainsky bullStreptomyces gliseus (Krainsky) Waksman and Henrici subspeciesstreptomycini (Krasilnikov) Lyons and Pridham strains with flexuouschains of spores spores smooth-walled yellow to yellowish-gray aeriamycelium occasionally tinged with pink yellow to yellowish-brownvegetative mycelium nonchromogenic utilizes D-glucose D-xyloseD-fructose D-galactose D-mannitol and possibly salicin but not Lshyarabinose L-rhamnose raffinose or i-inositol when incorporated in thePridham and Gottlieb chemically defined basal agar produces the


antibiotics streptomycin cycloheximide and an antibacterial antibiotic contained in the mycelium Holotype strain SAW 9 (3463) =ATCC 11429 one of the original streptomycin-producing strains isolated at Rutgers University

The status of the cultures studied that have red to purple vegetative growth is not clear at present There is little objective information on the identity of the antibiotics produced by these strains Until such information is obtained little can be said of their subspecific status Rhodomycetin and viomycins A B and C would seem to be antibiotics that should be considered as possible products of these strains

LITERATURE CITED (1) BENEDICT R G and LINDENFELSER L A

1951 PRODUCTION OF SOLUBLE PlGllENTS BY CERTAIN STRAINS OF STREPTDshyllYCES GRISEUS Antibiotics amp Chemotherapy 1 512-517

(2) BREED R S llURRAY E G and HITCHENS A P 1948 BERGEYS llANUAL OF DETERlllNATIVE B-ICTERIOLOGY Ed 6 948 pp

Baltimore Md (3) COllI111TTEE ON BACTERIOLOGICAL TECHNIC

1946 A ll-INUAL OF llETHODS FOR PURE CULTURE STUDY OF BACTERIA Geneva NY

(4) GORDON R E and Mum J M 1957 A COlIP-IRATIYE STUDY OF SOliE STRAINS RECEIVED AS NOCARDIA Jour

Bact 73 15-27 (5) -- and MIHll J M

1962 THE TYPE SPECIES OF THE GENUS NOCARDIA Jour Gen Microbiol 27 1-10

(6) --- and SmTH M M 1955 PROPOSED GROUP OF CHARACTERS FOR THE SEPAR-ITION OF STREPTOllYCES

AND NOCARDIA Jour Bact 69 147-150 (7) HESSELTINE C V BENEDICT R G and PRIDHUI T G

1954 USEFUL CRITERIA FOR SPECIES DIFFERENTIATION IN THE GENUS STREPTOshyMYCES N Y Acad Sci Ann 60 136-151

(8) INTERNATIONAL COlllllTTEE ON BACTERIOLOGICAL NOlIENCLATUHE EDITORIAL BOARD

1958INTERNATIONL CODE OF NOllECLATURE OF BACTERIA AND YIllUSESshyBACTERIOLOGICAL CODE HiS pp Ames Iowa

(9) KELlY K L and JUDD D B 1955 TilE ISCC-NBS llETIiOD OF DESIGNATING COLORS AND A DICTIONARY OF

COLOU NUIES US Dept Comm Cir 553 158 pp (10) KH-IINSKY A

1914 DIE AKTINOllYCETEX UND lHRE BEDEUTUNG IN DER NATUR Zentbl f Bakt Parasitenk Abt II) 41 649-688

(11) KUHERBERG A B STYLES H iINGHER H 0 and WAKSllAN S A 1950 THE PUODUCTION OF STREPTOCIN BY DIFFERENT STUAINS OF STREPTDshy

llYCES GRiSEUS Jour Bact 59 523-526 (12) KUSTEH R E

1961 RESULTS OF A COllPAHATIYE STUDY OF CRiTEHIA USED IN THE CLASSUICAshyTION OF THE ACTINOlIYCETES Internatl Bul Bact Nomencl Taxshyonomy 11 91-98

(13) LINDENFELSEU L A and PRIDHAll T G 1962 NATURAL VARIATIOX OF STREPTOlIYCES VIRIDOCHROllOGENES (KRAINSKY)

WAKSllAN AND HE~mICI Devlpmt and Indus Microbiol 3 245-256 (14) LYONS A J and PRIDHUI T G

1962 PROPOS-IL TO DESIGNATE STRAIX ATCC 3004 (IMRU 3(04) AS THE NEOTYPE STRAIN OF STREPTOlIYCES ALBUS (ROSSI-DORIJ) Jour Bact 83 370-380

(15) OKUII Y 1949 STUDIES ON THE ANTIBIOTIC STRAINS OF STREPTOllYCES II ON THE1--

STREPTOlIYCIN-PRODUCING STHAINS Jour Antibiotics (Tokyo) 2 Suppl A 85-87


(16) 1952 CLSSIFICATION O~ THE UiTGOXISTIC HAY FUNGI OF THE FAMILY

STREPTO~IYCETCEE OF HPAN Thesis Hokkaido University Japan 70 pp

(17) PRIDllUI T G 1965 COLOH AND STREPTmIYCETES REPORT OF N INTERNATIONAL WORKshy

SHOP ON DETER~IINATION OF COLOR OF STREPTOMYCETES Appl Microbiol 13 43-61

(18) --- and GOTTLIEB D 1948 THE UTILIZATION OF CARBON cmlPOUNDS BY smlE ACTINOMYCETALES

AS 11 ID FOR SPECIES DETER~IINTION Jour Bact 56 107-114 (19) --- HESSELTINE C V and BENEDICT R G

1958 GUIDE FOR TilE CLSSIFICTION OF STREPTOMYCETES CCOHDING TO SELECTED GROUPS PLCE~IENT OF STRAINS IS ~IORPHOLOGICAL SECTIONS Appl Microbiol 6 52-79

(20) --- LISDESFELSER L A SHOTWELL O L and others 1956NTIBIOTICS GAINST PLNT DISESE I LBORTORy ND GREENHOUSE

SURVEY Phytopathology 46 568-575 (21) REILLY H C H RRIS D A and VKS~IN S A

1947 AN CTINOPH GE FOR STREPTO~lyCES GRISEUS Jour Bact 54 451-466 (22) RIDGwYR

1912 COLOR STND RDS AND COLOR NOMESCLATURE Published by the author 43 pp 53 plates Washington DC

(23) ROBISON R S and NICKERSOS W J 1961 ENznl TIr DEH IRING OF HIDES SD SKINS US Patent No 2988488

(24) SHOCK~IN G and V KS~I N S A 1951 RHODO~IYCIN-N NTIBIOTIC PRODUCED By RED-PIGMENTED MUTNT

OF STREPTO~IYCES GRISEUS Antibiotics amp Chemotherapy 1 68-75 (25) S~IOLELIS A N and H RTSELL S E

1949 THE DETER~IINTIOS OF LYSOZnIE Jour Bact 58 731-736 (26) SUBco~mITTEE os TilE TAXONmlY OF THE ACTINmIYCETES

1958 ~IETIIODS FOR USE IN COOPERTIVE STLES ON CRITERI FOR DESCRIPshyTION OF TilE STREPTmIYCE BS 17 pp [Processed copies available from the Subcommittee Chairman Dr David Gottlieb University of Illinois Urbana1

(27) T YLOR H D KNOCHE L and GRNVILLE V C 1958 COLOR HR~IONY ~I NUL Ed 4 Container Corp of America

Chicago Ill (28) TRESNER H D and B CKUS E J

1963 SYSTE~I OF COLOR WHEELS FOR STREPTOMYCETE TXONOMY Appl Microbiol 11 335-338

(29) --- D VIES M C and B CKUS E J 1961 ELECTRON MICROSCOPY OF STREPTOMYCES SPORE MORPHOLOGY AND

ITS ROLE IN SPECIES DIFFEHENTITION Jour Bact 81 70-80 (30) U~IEZAW H OGAT Y TAKEUCHr T and T OT T

1948 ISOLTION OF STREPTOMYCIN FROI PINK VARINT OF STREPTOMYCES GRISEUS Japan Med Jour 1 397-404

(31) --- OGT Y T KEUCHr T and T BAT T 1949 ISOLTION OF STREPTO~IYCIN FROM PINK VARINrs OF STREPTOMYCES

GRISEUS Jour Antibiotics (Japan) 2 489-496 (32) WAKSMAN S A

1919 CULTURAL STUDIES OF SPECIES OF ACTINOMYCES Soil Sci 8 71-125 (33)

1959 STR IN SPECIFICITY ND PIIODUCTION OF NTIBIOTIC SUBSTANCES X CHARACTERIZATION AND CLASSIFICATION OF SPECIES WITHIN THE STREPT01IYCES GRISEUS GROUP Natl Acad Sci Proc 45 1043-1047

(34) --- and CURTIS R E 1916 THE CTINOMYCES OF TilE SOIL Soil Sci I 99-143

(35) --- and GREGORY F J 1954 ACTINOMYCIN II CLASSIFICATION OF ORGANISMS PRODUCING DIFFERENT

FOR~IS OF ACTINOMYCIN Antibiotics amp Chemotherapy 4 1050-1056 (36) --- H RIIIS D A KUPFERBERG A B and others

1949 STREPTOCIN ANTIBIOTIC ISOLATED FIIOll MYCELIUM OF STREPT01IYCES GRISEUS ACTIVE AGAINST TRICHOMONAS VAGINALIS AND CERTAIN BACTERIA Soc Expt BioI Med Proc 70 308-312


(37) --- REILLY H C and fuRRIS n A 1948 STREPTOMYCES GRISEUS (KRAINSKY) WAKBMAN AND HENRICI Jour

Bact 56 259-269 (38) WARREN H B JR PROKOP J F and GRUNDY W E

1955 NONSYNTHETIC MEDIA FOR ANTIBIOTIC-PRODUCING ACTINOMYCETES Antibiotics amp Chemotherapy 5 6-12

(39) WHIFFEN A J 1948 THE PRODUCTION ASSAY AND ANTIBIOTIC ACTIVITY OF ACTIDIONE AN

ANTIBIOTIC FIlOM STREPTOMYCES GRISEUS Jour Bact 56 283-291 (40) YAMAGUCHI T and SABURI Y

1955 STUDIES ON THE ANTI-TRICHOMONAL ACTINOMYCETES AND THEIR CLASSshyIFICATION Jour Gen Appl Microbiol 1 201-235

bull

VS GCMINMDIT PIIINTII IIffICt l1li-0 II

f

tmiddot

REFtCNCE DO NOT LOAN



A Taxonomic Study f Some Strains

Technical Bulletin No 1360

-

F--shy

ifshy~o

0 ~

~

Agricultural Research Service United States Department of Agriculture





li

_________________________________________________ _


P~oteolytlC a~tVlty





fge 1 1 6 7 7 7 7 7 8 8 8 8 8 8 9 9

10 10 10 11 11 11 11 13 13 14 15 15 16 16 17 17 19 25 29


iii














1




activity



B-128L________ grisells_________ (1)


IPV 423x_______ griseu8_________ (I)

NCIB 8225 _____ griseuL________ (1)














activity

NCIB 9004 _____

S-1471- ________

SAW 4-L ______

SAW 4-2 ______

SAW 4-3 _______

SAW 4-4 _______

SAW 4-5 _______

SAW 4-6 _______

SAW 4-7 _______

SAW 9-L ______

griseus _________

griseuB- ________

griseus _________

griseus _________

griseus _________

griseuB- _______

griseus _________

griseu8 _________

griseus _________

griseus _________

B12 ___ -- _ --- ---- shy

(I)



streptomycin______

streptomycin______

streptomycin______

streptomycin______

(1)

streptomycin______















activity

















________________

________________



Designation used

NIHT SN-2 (2) __

NIHJ SN-14-L_

NIHJ SN-14-2__

Lilly L _________


ARI1780_______

NI9003 ________

Carpenter_______

E______________

Specific epithet




sp _____________

sp _____________ sp _____________ sp _____________ sp _____________

streptomycini____

vinaceus________


activity

streptomycin______

streptomycin______

streptomycin______

streptomycin_____ _


complex

(1)

(1)

(1)

(1)






Do Do Do







----------------

----------------




activity







---------------




--------------shy




---------------shy

8-1765 _________ ________________ (1) Do8-1766 _________________________ (I) Do8-1767 _________________________ (1) Do






Diastotic Activity








Color






























RESULTS








Diastatic Activity



Nitrate Reduction






Designation used


S-62____________________________ _~~~e~~~~=======================8-901___________________________ _ S-1757__________________________ _ 8-1758__________________________ _8-1759__________________________ _ 8-1760__________________________ _ 8-1761__________________________ _8-1762__________________________ _ 8-1763__________________________ _ 8-1764__________________________ _ 8-1765 __________________________ _ 8-1766__________________________ _8-1767__________________________ _


Tube 1 Tube 2

+ + C+) (+)(+) (+) C+) C+)

(-)

C+) (-)C+) (-)

(+) (+)

(-) (-) (-) (-)

C+) (+)

(+) (+)

(-) (-) (+) (+)

(-)

(-) (-)

(-) (-)

(+) (+)(+) C+)(- ) (- )

C+) (- )(+) (-) (- ) C+) C+)(-) (- )

C+) (+) (- ) (- )

(- )

c+) (+) + +

Organic-based broth

Tube 1 Tube 2

(+) + +

(+) (+) + + - -

(+) (+)c+) (+)

(+) + +

c+) C+)(- ) c+) (+)c-) C-)c+) C+)(- ) c-)(- )

+ +(+) (- ) + + - (+)

(+) + + + + -

(- ) (-) (- )

(- ) (-) (- ) C+) C+)(+) + -C+) C+)

c+) (+) + + + +

C+) c+)c+) c+) + +

(+) c+)

c+) C+)(- ) c-) + + + +

(+) (+)C+) (+) + + + +



- -


B

(p S D


















Tresner name and

color wheel

Pridhllm color

NBS color

NBS color


desigshynation 4

series (28)

series (19)

names of Ridgway

tabs 5

names of CUM tabs 5

















_____

________













































Red 262












01 gt l Il0 a 01 01 0






(- ) + + - + - c+)


Carpenter _________________ - + + (- ) - + + - + - (+)- + + - - + + - + - (- )

~------------------------_____________________ + - + - + (+)S-62 - + - + shy(- ) shyS-901 ____________________ - + + - + + + - (- ) (-) shy8-1757 ___________________ - + + - + + + - (- ) - Ii8-1758 ___________________ - + + - + + - + - c-)

S-1759 ___________________ - + + (- ) - + + - + - c-) 8-1760 ___________________ - + + - - + + - + - c-) 8-1761 ___________________ - + C+) - - + + - + - c+) 8-1762 ___________________ - + C+) - - (+) + - + - (+) 8-1763 ___________ - _______ - + + - - + + - + - c-) S-1764 ___________________ - + + - + + - + - (+)

- + + - + + - + - c+)C==)18-1765_ - - bull -___________________ - - - - - - __ - - ---OJ + - + + - (+)8-1766 - + + shy8-1767 ___________________ - + + - + + - + - C+)

- + - - (+) + - (+) - (- )I+ I shy


------------------





Tube 1 Tube 2










cent 22 22 18 22 23 19 20

cent 50 72 34 39 36 30 57

cent 66 81 51 61 62 49 64

cent 73 82 66 73 76 67 70

cent 75 83 76 76 75 77 73

cent 76 88 81 80 76 84 77

cent 0 85 82 81 81 87 80


20 ca 95 96 96 96 96 96


Antibiotic Activi~y







medium 2










Designation used

01 01 1




I I i1





- -







N - ~~ - ~~Czc ~z CQ UJ



l







ci 00 ~



~l il ~l ~l lt~ sectl~~ ~~ g~ ~ ~~ ~~ ~ ~~ 6~~~

~~






I 0




ci 00 ~ g 5 g Cj r- ~~Ogt 0 ~r-




0 0 0

21 0 0

32 0 0

15 1lt 0

0 0 0

0 0 0


0 0 0

18 0 0

26 0 0

(3) 0 0

0 0 0

0 0 0


0 0 0

20 0 0

32 0 0

16 14 0

0 0 0

0 0 0


0 0 0

22 0 0

35 0 0

19 0 0

0 0 0

0 0



0 0

18 18

0 0

0 0

0 0

0 0


0 0

15 14

0 0

(3) (3)

0 0

0 0





-- -- ---- ----











---- 5 7 7 [) 3 0 0 3 4 0









Oritil





Orilin







Orilil





bull

WSI ISW 10 NHCI







































































bull


f

tmiddot





li

_________________________________________________ _


P~oteolytlC a~tVlty





fge 1 1 6 7 7 7 7 7 8 8 8 8 8 8 9 9

10 10 10 11 11 11 11 13 13 14 15 15 16 16 17 17 19 25 29


iii














1




activity



B-128L________ grisells_________ (1)


IPV 423x_______ griseu8_________ (I)

NCIB 8225 _____ griseuL________ (1)














activity

NCIB 9004 _____

S-1471- ________

SAW 4-L ______

SAW 4-2 ______

SAW 4-3 _______

SAW 4-4 _______

SAW 4-5 _______

SAW 4-6 _______

SAW 4-7 _______

SAW 9-L ______

griseus _________

griseuB- ________

griseus _________

griseus _________

griseus _________

griseuB- _______

griseus _________

griseu8 _________

griseus _________

griseus _________

B12 ___ -- _ --- ---- shy

(I)



streptomycin______

streptomycin______

streptomycin______

streptomycin______

(1)

streptomycin______















activity

















________________

________________



Designation used

NIHT SN-2 (2) __

NIHJ SN-14-L_

NIHJ SN-14-2__

Lilly L _________


ARI1780_______

NI9003 ________

Carpenter_______

E______________

Specific epithet




sp _____________

sp _____________ sp _____________ sp _____________ sp _____________

streptomycini____

vinaceus________


activity

streptomycin______

streptomycin______

streptomycin______

streptomycin_____ _


complex

(1)

(1)

(1)

(1)






Do Do Do







----------------

----------------




activity







---------------




--------------shy




---------------shy

8-1765 _________ ________________ (1) Do8-1766 _________________________ (I) Do8-1767 _________________________ (1) Do






Diastotic Activity








Color






























RESULTS








Diastatic Activity



Nitrate Reduction






Designation used


S-62____________________________ _~~~e~~~~=======================8-901___________________________ _ S-1757__________________________ _ 8-1758__________________________ _8-1759__________________________ _ 8-1760__________________________ _ 8-1761__________________________ _8-1762__________________________ _ 8-1763__________________________ _ 8-1764__________________________ _ 8-1765 __________________________ _ 8-1766__________________________ _8-1767__________________________ _


Tube 1 Tube 2

+ + C+) (+)(+) (+) C+) C+)

(-)

C+) (-)C+) (-)

(+) (+)

(-) (-) (-) (-)

C+) (+)

(+) (+)

(-) (-) (+) (+)

(-)

(-) (-)

(-) (-)

(+) (+)(+) C+)(- ) (- )

C+) (- )(+) (-) (- ) C+) C+)(-) (- )

C+) (+) (- ) (- )

(- )

c+) (+) + +

Organic-based broth

Tube 1 Tube 2

(+) + +

(+) (+) + + - -

(+) (+)c+) (+)

(+) + +

c+) C+)(- ) c+) (+)c-) C-)c+) C+)(- ) c-)(- )

+ +(+) (- ) + + - (+)

(+) + + + + -

(- ) (-) (- )

(- ) (-) (- ) C+) C+)(+) + -C+) C+)

c+) (+) + + + +

C+) c+)c+) c+) + +

(+) c+)

c+) C+)(- ) c-) + + + +

(+) (+)C+) (+) + + + +



- -


B

(p S D


















Tresner name and

color wheel

Pridhllm color

NBS color

NBS color


desigshynation 4

series (28)

series (19)

names of Ridgway

tabs 5

names of CUM tabs 5

















_____

________













































Red 262












01 gt l Il0 a 01 01 0






(- ) + + - + - c+)


Carpenter _________________ - + + (- ) - + + - + - (+)- + + - - + + - + - (- )

~------------------------_____________________ + - + - + (+)S-62 - + - + shy(- ) shyS-901 ____________________ - + + - + + + - (- ) (-) shy8-1757 ___________________ - + + - + + + - (- ) - Ii8-1758 ___________________ - + + - + + - + - c-)

S-1759 ___________________ - + + (- ) - + + - + - c-) 8-1760 ___________________ - + + - - + + - + - c-) 8-1761 ___________________ - + C+) - - + + - + - c+) 8-1762 ___________________ - + C+) - - (+) + - + - (+) 8-1763 ___________ - _______ - + + - - + + - + - c-) S-1764 ___________________ - + + - + + - + - (+)

- + + - + + - + - c+)C==)18-1765_ - - bull -___________________ - - - - - - __ - - ---OJ + - + + - (+)8-1766 - + + shy8-1767 ___________________ - + + - + + - + - C+)

- + - - (+) + - (+) - (- )I+ I shy


------------------





Tube 1 Tube 2










cent 22 22 18 22 23 19 20

cent 50 72 34 39 36 30 57

cent 66 81 51 61 62 49 64

cent 73 82 66 73 76 67 70

cent 75 83 76 76 75 77 73

cent 76 88 81 80 76 84 77

cent 0 85 82 81 81 87 80


20 ca 95 96 96 96 96 96


Antibiotic Activi~y







medium 2










Designation used

01 01 1




I I i1





- -







N - ~~ - ~~Czc ~z CQ UJ



l







ci 00 ~



~l il ~l ~l lt~ sectl~~ ~~ g~ ~ ~~ ~~ ~ ~~ 6~~~

~~






I 0




ci 00 ~ g 5 g Cj r- ~~Ogt 0 ~r-




0 0 0

21 0 0

32 0 0

15 1lt 0

0 0 0

0 0 0


0 0 0

18 0 0

26 0 0

(3) 0 0

0 0 0

0 0 0


0 0 0

20 0 0

32 0 0

16 14 0

0 0 0

0 0 0


0 0 0

22 0 0

35 0 0

19 0 0

0 0 0

0 0



0 0

18 18

0 0

0 0

0 0

0 0


0 0

15 14

0 0

(3) (3)

0 0

0 0





-- -- ---- ----











---- 5 7 7 [) 3 0 0 3 4 0









Oritil





Orilin







Orilil





bull

WSI ISW 10 NHCI







































































bull


f

tmiddot

_________________________________________________ _


P~oteolytlC a~tVlty





fge 1 1 6 7 7 7 7 7 8 8 8 8 8 8 9 9

10 10 10 11 11 11 11 13 13 14 15 15 16 16 17 17 19 25 29


iii














1




activity



B-128L________ grisells_________ (1)


IPV 423x_______ griseu8_________ (I)

NCIB 8225 _____ griseuL________ (1)














activity

NCIB 9004 _____

S-1471- ________

SAW 4-L ______

SAW 4-2 ______

SAW 4-3 _______

SAW 4-4 _______

SAW 4-5 _______

SAW 4-6 _______

SAW 4-7 _______

SAW 9-L ______

griseus _________

griseuB- ________

griseus _________

griseus _________

griseus _________

griseuB- _______

griseus _________

griseu8 _________

griseus _________

griseus _________

B12 ___ -- _ --- ---- shy

(I)



streptomycin______

streptomycin______

streptomycin______

streptomycin______

(1)

streptomycin______















activity

















________________

________________



Designation used

NIHT SN-2 (2) __

NIHJ SN-14-L_

NIHJ SN-14-2__

Lilly L _________


ARI1780_______

NI9003 ________

Carpenter_______

E______________

Specific epithet




sp _____________

sp _____________ sp _____________ sp _____________ sp _____________

streptomycini____

vinaceus________


activity

streptomycin______

streptomycin______

streptomycin______

streptomycin_____ _


complex

(1)

(1)

(1)

(1)






Do Do Do







----------------

----------------




activity







---------------




--------------shy




---------------shy

8-1765 _________ ________________ (1) Do8-1766 _________________________ (I) Do8-1767 _________________________ (1) Do






Diastotic Activity








Color






























RESULTS








Diastatic Activity



Nitrate Reduction






Designation used


S-62____________________________ _~~~e~~~~=======================8-901___________________________ _ S-1757__________________________ _ 8-1758__________________________ _8-1759__________________________ _ 8-1760__________________________ _ 8-1761__________________________ _8-1762__________________________ _ 8-1763__________________________ _ 8-1764__________________________ _ 8-1765 __________________________ _ 8-1766__________________________ _8-1767__________________________ _


Tube 1 Tube 2

+ + C+) (+)(+) (+) C+) C+)

(-)

C+) (-)C+) (-)

(+) (+)

(-) (-) (-) (-)

C+) (+)

(+) (+)

(-) (-) (+) (+)

(-)

(-) (-)

(-) (-)

(+) (+)(+) C+)(- ) (- )

C+) (- )(+) (-) (- ) C+) C+)(-) (- )

C+) (+) (- ) (- )

(- )

c+) (+) + +

Organic-based broth

Tube 1 Tube 2

(+) + +

(+) (+) + + - -

(+) (+)c+) (+)

(+) + +

c+) C+)(- ) c+) (+)c-) C-)c+) C+)(- ) c-)(- )

+ +(+) (- ) + + - (+)

(+) + + + + -

(- ) (-) (- )

(- ) (-) (- ) C+) C+)(+) + -C+) C+)

c+) (+) + + + +

C+) c+)c+) c+) + +

(+) c+)

c+) C+)(- ) c-) + + + +

(+) (+)C+) (+) + + + +



- -


B

(p S D


















Tresner name and

color wheel

Pridhllm color

NBS color

NBS color


desigshynation 4

series (28)

series (19)

names of Ridgway

tabs 5

names of CUM tabs 5

















_____

________













































Red 262












01 gt l Il0 a 01 01 0






(- ) + + - + - c+)


Carpenter _________________ - + + (- ) - + + - + - (+)- + + - - + + - + - (- )

~------------------------_____________________ + - + - + (+)S-62 - + - + shy(- ) shyS-901 ____________________ - + + - + + + - (- ) (-) shy8-1757 ___________________ - + + - + + + - (- ) - Ii8-1758 ___________________ - + + - + + - + - c-)

S-1759 ___________________ - + + (- ) - + + - + - c-) 8-1760 ___________________ - + + - - + + - + - c-) 8-1761 ___________________ - + C+) - - + + - + - c+) 8-1762 ___________________ - + C+) - - (+) + - + - (+) 8-1763 ___________ - _______ - + + - - + + - + - c-) S-1764 ___________________ - + + - + + - + - (+)

- + + - + + - + - c+)C==)18-1765_ - - bull -___________________ - - - - - - __ - - ---OJ + - + + - (+)8-1766 - + + shy8-1767 ___________________ - + + - + + - + - C+)

- + - - (+) + - (+) - (- )I+ I shy


------------------





Tube 1 Tube 2










cent 22 22 18 22 23 19 20

cent 50 72 34 39 36 30 57

cent 66 81 51 61 62 49 64

cent 73 82 66 73 76 67 70

cent 75 83 76 76 75 77 73

cent 76 88 81 80 76 84 77

cent 0 85 82 81 81 87 80


20 ca 95 96 96 96 96 96


Antibiotic Activi~y







medium 2










Designation used

01 01 1




I I i1





- -







N - ~~ - ~~Czc ~z CQ UJ



l







ci 00 ~



~l il ~l ~l lt~ sectl~~ ~~ g~ ~ ~~ ~~ ~ ~~ 6~~~

~~






I 0




ci 00 ~ g 5 g Cj r- ~~Ogt 0 ~r-




0 0 0

21 0 0

32 0 0

15 1lt 0

0 0 0

0 0 0


0 0 0

18 0 0

26 0 0

(3) 0 0

0 0 0

0 0 0


0 0 0

20 0 0

32 0 0

16 14 0

0 0 0

0 0 0


0 0 0

22 0 0

35 0 0

19 0 0

0 0 0

0 0



0 0

18 18

0 0

0 0

0 0

0 0


0 0

15 14

0 0

(3) (3)

0 0

0 0





-- -- ---- ----











---- 5 7 7 [) 3 0 0 3 4 0









Oritil





Orilin







Orilil





bull

WSI ISW 10 NHCI







































































bull


f

tmiddot














1




activity



B-128L________ grisells_________ (1)


IPV 423x_______ griseu8_________ (I)

NCIB 8225 _____ griseuL________ (1)














activity

NCIB 9004 _____

S-1471- ________

SAW 4-L ______

SAW 4-2 ______

SAW 4-3 _______

SAW 4-4 _______

SAW 4-5 _______

SAW 4-6 _______

SAW 4-7 _______

SAW 9-L ______

griseus _________

griseuB- ________

griseus _________

griseus _________

griseus _________

griseuB- _______

griseus _________

griseu8 _________

griseus _________

griseus _________

B12 ___ -- _ --- ---- shy

(I)



streptomycin______

streptomycin______

streptomycin______

streptomycin______

(1)

streptomycin______















activity

















________________

________________



Designation used

NIHT SN-2 (2) __

NIHJ SN-14-L_

NIHJ SN-14-2__

Lilly L _________


ARI1780_______

NI9003 ________

Carpenter_______

E______________

Specific epithet




sp _____________

sp _____________ sp _____________ sp _____________ sp _____________

streptomycini____

vinaceus________


activity

streptomycin______

streptomycin______

streptomycin______

streptomycin_____ _


complex

(1)

(1)

(1)

(1)






Do Do Do







----------------

----------------




activity







---------------




--------------shy




---------------shy

8-1765 _________ ________________ (1) Do8-1766 _________________________ (I) Do8-1767 _________________________ (1) Do






Diastotic Activity








Color






























RESULTS








Diastatic Activity



Nitrate Reduction






Designation used


S-62____________________________ _~~~e~~~~=======================8-901___________________________ _ S-1757__________________________ _ 8-1758__________________________ _8-1759__________________________ _ 8-1760__________________________ _ 8-1761__________________________ _8-1762__________________________ _ 8-1763__________________________ _ 8-1764__________________________ _ 8-1765 __________________________ _ 8-1766__________________________ _8-1767__________________________ _


Tube 1 Tube 2

+ + C+) (+)(+) (+) C+) C+)

(-)

C+) (-)C+) (-)

(+) (+)

(-) (-) (-) (-)

C+) (+)

(+) (+)

(-) (-) (+) (+)

(-)

(-) (-)

(-) (-)

(+) (+)(+) C+)(- ) (- )

C+) (- )(+) (-) (- ) C+) C+)(-) (- )

C+) (+) (- ) (- )

(- )

c+) (+) + +

Organic-based broth

Tube 1 Tube 2

(+) + +

(+) (+) + + - -

(+) (+)c+) (+)

(+) + +

c+) C+)(- ) c+) (+)c-) C-)c+) C+)(- ) c-)(- )

+ +(+) (- ) + + - (+)

(+) + + + + -

(- ) (-) (- )

(- ) (-) (- ) C+) C+)(+) + -C+) C+)

c+) (+) + + + +

C+) c+)c+) c+) + +

(+) c+)

c+) C+)(- ) c-) + + + +

(+) (+)C+) (+) + + + +



- -


B

(p S D


















Tresner name and

color wheel

Pridhllm color

NBS color

NBS color


desigshynation 4

series (28)

series (19)

names of Ridgway

tabs 5

names of CUM tabs 5

















_____

________













































Red 262












01 gt l Il0 a 01 01 0






(- ) + + - + - c+)


Carpenter _________________ - + + (- ) - + + - + - (+)- + + - - + + - + - (- )

~------------------------_____________________ + - + - + (+)S-62 - + - + shy(- ) shyS-901 ____________________ - + + - + + + - (- ) (-) shy8-1757 ___________________ - + + - + + + - (- ) - Ii8-1758 ___________________ - + + - + + - + - c-)

S-1759 ___________________ - + + (- ) - + + - + - c-) 8-1760 ___________________ - + + - - + + - + - c-) 8-1761 ___________________ - + C+) - - + + - + - c+) 8-1762 ___________________ - + C+) - - (+) + - + - (+) 8-1763 ___________ - _______ - + + - - + + - + - c-) S-1764 ___________________ - + + - + + - + - (+)

- + + - + + - + - c+)C==)18-1765_ - - bull -___________________ - - - - - - __ - - ---OJ + - + + - (+)8-1766 - + + shy8-1767 ___________________ - + + - + + - + - C+)

- + - - (+) + - (+) - (- )I+ I shy


------------------





Tube 1 Tube 2










cent 22 22 18 22 23 19 20

cent 50 72 34 39 36 30 57

cent 66 81 51 61 62 49 64

cent 73 82 66 73 76 67 70

cent 75 83 76 76 75 77 73

cent 76 88 81 80 76 84 77

cent 0 85 82 81 81 87 80


20 ca 95 96 96 96 96 96


Antibiotic Activi~y







medium 2










Designation used

01 01 1




I I i1





- -







N - ~~ - ~~Czc ~z CQ UJ



l







ci 00 ~



~l il ~l ~l lt~ sectl~~ ~~ g~ ~ ~~ ~~ ~ ~~ 6~~~

~~






I 0




ci 00 ~ g 5 g Cj r- ~~Ogt 0 ~r-




0 0 0

21 0 0

32 0 0

15 1lt 0

0 0 0

0 0 0


0 0 0

18 0 0

26 0 0

(3) 0 0

0 0 0

0 0 0


0 0 0

20 0 0

32 0 0

16 14 0

0 0 0

0 0 0


0 0 0

22 0 0

35 0 0

19 0 0

0 0 0

0 0



0 0

18 18

0 0

0 0

0 0

0 0


0 0

15 14

0 0

(3) (3)

0 0

0 0





-- -- ---- ----











---- 5 7 7 [) 3 0 0 3 4 0









Oritil





Orilin







Orilil





bull

WSI ISW 10 NHCI







































































bull


f

tmiddot




activity



B-128L________ grisells_________ (1)


IPV 423x_______ griseu8_________ (I)

NCIB 8225 _____ griseuL________ (1)














activity

NCIB 9004 _____

S-1471- ________

SAW 4-L ______

SAW 4-2 ______

SAW 4-3 _______

SAW 4-4 _______

SAW 4-5 _______

SAW 4-6 _______

SAW 4-7 _______

SAW 9-L ______

griseus _________

griseuB- ________

griseus _________

griseus _________

griseus _________

griseuB- _______

griseus _________

griseu8 _________

griseus _________

griseus _________

B12 ___ -- _ --- ---- shy

(I)



streptomycin______

streptomycin______

streptomycin______

streptomycin______

(1)

streptomycin______















activity

















________________

________________



Designation used

NIHT SN-2 (2) __

NIHJ SN-14-L_

NIHJ SN-14-2__

Lilly L _________


ARI1780_______

NI9003 ________

Carpenter_______

E______________

Specific epithet




sp _____________

sp _____________ sp _____________ sp _____________ sp _____________

streptomycini____

vinaceus________


activity

streptomycin______

streptomycin______

streptomycin______

streptomycin_____ _


complex

(1)

(1)

(1)

(1)






Do Do Do







----------------

----------------




activity







---------------




--------------shy




---------------shy

8-1765 _________ ________________ (1) Do8-1766 _________________________ (I) Do8-1767 _________________________ (1) Do






Diastotic Activity








Color






























RESULTS








Diastatic Activity



Nitrate Reduction






Designation used


S-62____________________________ _~~~e~~~~=======================8-901___________________________ _ S-1757__________________________ _ 8-1758__________________________ _8-1759__________________________ _ 8-1760__________________________ _ 8-1761__________________________ _8-1762__________________________ _ 8-1763__________________________ _ 8-1764__________________________ _ 8-1765 __________________________ _ 8-1766__________________________ _8-1767__________________________ _


Tube 1 Tube 2

+ + C+) (+)(+) (+) C+) C+)

(-)

C+) (-)C+) (-)

(+) (+)

(-) (-) (-) (-)

C+) (+)

(+) (+)

(-) (-) (+) (+)

(-)

(-) (-)

(-) (-)

(+) (+)(+) C+)(- ) (- )

C+) (- )(+) (-) (- ) C+) C+)(-) (- )

C+) (+) (- ) (- )

(- )

c+) (+) + +

Organic-based broth

Tube 1 Tube 2

(+) + +

(+) (+) + + - -

(+) (+)c+) (+)

(+) + +

c+) C+)(- ) c+) (+)c-) C-)c+) C+)(- ) c-)(- )

+ +(+) (- ) + + - (+)

(+) + + + + -

(- ) (-) (- )

(- ) (-) (- ) C+) C+)(+) + -C+) C+)

c+) (+) + + + +

C+) c+)c+) c+) + +

(+) c+)

c+) C+)(- ) c-) + + + +

(+) (+)C+) (+) + + + +



- -


B

(p S D


















Tresner name and

color wheel

Pridhllm color

NBS color

NBS color


desigshynation 4

series (28)

series (19)

names of Ridgway

tabs 5

names of CUM tabs 5

















_____

________













































Red 262












01 gt l Il0 a 01 01 0






(- ) + + - + - c+)


Carpenter _________________ - + + (- ) - + + - + - (+)- + + - - + + - + - (- )

~------------------------_____________________ + - + - + (+)S-62 - + - + shy(- ) shyS-901 ____________________ - + + - + + + - (- ) (-) shy8-1757 ___________________ - + + - + + + - (- ) - Ii8-1758 ___________________ - + + - + + - + - c-)

S-1759 ___________________ - + + (- ) - + + - + - c-) 8-1760 ___________________ - + + - - + + - + - c-) 8-1761 ___________________ - + C+) - - + + - + - c+) 8-1762 ___________________ - + C+) - - (+) + - + - (+) 8-1763 ___________ - _______ - + + - - + + - + - c-) S-1764 ___________________ - + + - + + - + - (+)

- + + - + + - + - c+)C==)18-1765_ - - bull -___________________ - - - - - - __ - - ---OJ + - + + - (+)8-1766 - + + shy8-1767 ___________________ - + + - + + - + - C+)

- + - - (+) + - (+) - (- )I+ I shy


------------------





Tube 1 Tube 2










cent 22 22 18 22 23 19 20

cent 50 72 34 39 36 30 57

cent 66 81 51 61 62 49 64

cent 73 82 66 73 76 67 70

cent 75 83 76 76 75 77 73

cent 76 88 81 80 76 84 77

cent 0 85 82 81 81 87 80


20 ca 95 96 96 96 96 96


Antibiotic Activi~y







medium 2










Designation used

01 01 1




I I i1





- -







N - ~~ - ~~Czc ~z CQ UJ



l







ci 00 ~



~l il ~l ~l lt~ sectl~~ ~~ g~ ~ ~~ ~~ ~ ~~ 6~~~

~~






I 0




ci 00 ~ g 5 g Cj r- ~~Ogt 0 ~r-




0 0 0

21 0 0

32 0 0

15 1lt 0

0 0 0

0 0 0


0 0 0

18 0 0

26 0 0

(3) 0 0

0 0 0

0 0 0


0 0 0

20 0 0

32 0 0

16 14 0

0 0 0

0 0 0


0 0 0

22 0 0

35 0 0

19 0 0

0 0 0

0 0



0 0

18 18

0 0

0 0

0 0

0 0


0 0

15 14

0 0

(3) (3)

0 0

0 0





-- -- ---- ----











---- 5 7 7 [) 3 0 0 3 4 0









Oritil





Orilin







Orilil





bull

WSI ISW 10 NHCI







































































bull


f

tmiddot




activity

NCIB 9004 _____

S-1471- ________

SAW 4-L ______

SAW 4-2 ______

SAW 4-3 _______

SAW 4-4 _______

SAW 4-5 _______

SAW 4-6 _______

SAW 4-7 _______

SAW 9-L ______

griseus _________

griseuB- ________

griseus _________

griseus _________

griseus _________

griseuB- _______

griseus _________

griseu8 _________

griseus _________

griseus _________

B12 ___ -- _ --- ---- shy

(I)



streptomycin______

streptomycin______

streptomycin______

streptomycin______

(1)

streptomycin______















activity

















________________

________________



Designation used

NIHT SN-2 (2) __

NIHJ SN-14-L_

NIHJ SN-14-2__

Lilly L _________


ARI1780_______

NI9003 ________

Carpenter_______

E______________

Specific epithet




sp _____________

sp _____________ sp _____________ sp _____________ sp _____________

streptomycini____

vinaceus________


activity

streptomycin______

streptomycin______

streptomycin______

streptomycin_____ _


complex

(1)

(1)

(1)

(1)






Do Do Do







----------------

----------------




activity







---------------




--------------shy




---------------shy

8-1765 _________ ________________ (1) Do8-1766 _________________________ (I) Do8-1767 _________________________ (1) Do






Diastotic Activity








Color






























RESULTS








Diastatic Activity



Nitrate Reduction






Designation used


S-62____________________________ _~~~e~~~~=======================8-901___________________________ _ S-1757__________________________ _ 8-1758__________________________ _8-1759__________________________ _ 8-1760__________________________ _ 8-1761__________________________ _8-1762__________________________ _ 8-1763__________________________ _ 8-1764__________________________ _ 8-1765 __________________________ _ 8-1766__________________________ _8-1767__________________________ _


Tube 1 Tube 2

+ + C+) (+)(+) (+) C+) C+)

(-)

C+) (-)C+) (-)

(+) (+)

(-) (-) (-) (-)

C+) (+)

(+) (+)

(-) (-) (+) (+)

(-)

(-) (-)

(-) (-)

(+) (+)(+) C+)(- ) (- )

C+) (- )(+) (-) (- ) C+) C+)(-) (- )

C+) (+) (- ) (- )

(- )

c+) (+) + +

Organic-based broth

Tube 1 Tube 2

(+) + +

(+) (+) + + - -

(+) (+)c+) (+)

(+) + +

c+) C+)(- ) c+) (+)c-) C-)c+) C+)(- ) c-)(- )

+ +(+) (- ) + + - (+)

(+) + + + + -

(- ) (-) (- )

(- ) (-) (- ) C+) C+)(+) + -C+) C+)

c+) (+) + + + +

C+) c+)c+) c+) + +

(+) c+)

c+) C+)(- ) c-) + + + +

(+) (+)C+) (+) + + + +



- -


B

(p S D


















Tresner name and

color wheel

Pridhllm color

NBS color

NBS color


desigshynation 4

series (28)

series (19)

names of Ridgway

tabs 5

names of CUM tabs 5

















_____

________













































Red 262












01 gt l Il0 a 01 01 0






(- ) + + - + - c+)


Carpenter _________________ - + + (- ) - + + - + - (+)- + + - - + + - + - (- )

~------------------------_____________________ + - + - + (+)S-62 - + - + shy(- ) shyS-901 ____________________ - + + - + + + - (- ) (-) shy8-1757 ___________________ - + + - + + + - (- ) - Ii8-1758 ___________________ - + + - + + - + - c-)

S-1759 ___________________ - + + (- ) - + + - + - c-) 8-1760 ___________________ - + + - - + + - + - c-) 8-1761 ___________________ - + C+) - - + + - + - c+) 8-1762 ___________________ - + C+) - - (+) + - + - (+) 8-1763 ___________ - _______ - + + - - + + - + - c-) S-1764 ___________________ - + + - + + - + - (+)

- + + - + + - + - c+)C==)18-1765_ - - bull -___________________ - - - - - - __ - - ---OJ + - + + - (+)8-1766 - + + shy8-1767 ___________________ - + + - + + - + - C+)

- + - - (+) + - (+) - (- )I+ I shy


------------------





Tube 1 Tube 2










cent 22 22 18 22 23 19 20

cent 50 72 34 39 36 30 57

cent 66 81 51 61 62 49 64

cent 73 82 66 73 76 67 70

cent 75 83 76 76 75 77 73

cent 76 88 81 80 76 84 77

cent 0 85 82 81 81 87 80


20 ca 95 96 96 96 96 96


Antibiotic Activi~y







medium 2










Designation used

01 01 1




I I i1





- -







N - ~~ - ~~Czc ~z CQ UJ



l







ci 00 ~



~l il ~l ~l lt~ sectl~~ ~~ g~ ~ ~~ ~~ ~ ~~ 6~~~

~~






I 0




ci 00 ~ g 5 g Cj r- ~~Ogt 0 ~r-




0 0 0

21 0 0

32 0 0

15 1lt 0

0 0 0

0 0 0


0 0 0

18 0 0

26 0 0

(3) 0 0

0 0 0

0 0 0


0 0 0

20 0 0

32 0 0

16 14 0

0 0 0

0 0 0


0 0 0

22 0 0

35 0 0

19 0 0

0 0 0

0 0



0 0

18 18

0 0

0 0

0 0

0 0


0 0

15 14

0 0

(3) (3)

0 0

0 0





-- -- ---- ----











---- 5 7 7 [) 3 0 0 3 4 0









Oritil





Orilin







Orilil





bull

WSI ISW 10 NHCI







































































bull


f

tmiddot




activity

















________________

________________



Designation used

NIHT SN-2 (2) __

NIHJ SN-14-L_

NIHJ SN-14-2__

Lilly L _________


ARI1780_______

NI9003 ________

Carpenter_______

E______________

Specific epithet




sp _____________

sp _____________ sp _____________ sp _____________ sp _____________

streptomycini____

vinaceus________


activity

streptomycin______

streptomycin______

streptomycin______

streptomycin_____ _


complex

(1)

(1)

(1)

(1)






Do Do Do







----------------

----------------




activity







---------------




--------------shy




---------------shy

8-1765 _________ ________________ (1) Do8-1766 _________________________ (I) Do8-1767 _________________________ (1) Do






Diastotic Activity








Color






























RESULTS








Diastatic Activity



Nitrate Reduction






Designation used


S-62____________________________ _~~~e~~~~=======================8-901___________________________ _ S-1757__________________________ _ 8-1758__________________________ _8-1759__________________________ _ 8-1760__________________________ _ 8-1761__________________________ _8-1762__________________________ _ 8-1763__________________________ _ 8-1764__________________________ _ 8-1765 __________________________ _ 8-1766__________________________ _8-1767__________________________ _


Tube 1 Tube 2

+ + C+) (+)(+) (+) C+) C+)

(-)

C+) (-)C+) (-)

(+) (+)

(-) (-) (-) (-)

C+) (+)

(+) (+)

(-) (-) (+) (+)

(-)

(-) (-)

(-) (-)

(+) (+)(+) C+)(- ) (- )

C+) (- )(+) (-) (- ) C+) C+)(-) (- )

C+) (+) (- ) (- )

(- )

c+) (+) + +

Organic-based broth

Tube 1 Tube 2

(+) + +

(+) (+) + + - -

(+) (+)c+) (+)

(+) + +

c+) C+)(- ) c+) (+)c-) C-)c+) C+)(- ) c-)(- )

+ +(+) (- ) + + - (+)

(+) + + + + -

(- ) (-) (- )

(- ) (-) (- ) C+) C+)(+) + -C+) C+)

c+) (+) + + + +

C+) c+)c+) c+) + +

(+) c+)

c+) C+)(- ) c-) + + + +

(+) (+)C+) (+) + + + +



- -


B

(p S D


















Tresner name and

color wheel

Pridhllm color

NBS color

NBS color


desigshynation 4

series (28)

series (19)

names of Ridgway

tabs 5

names of CUM tabs 5

















_____

________













































Red 262












01 gt l Il0 a 01 01 0






(- ) + + - + - c+)


Carpenter _________________ - + + (- ) - + + - + - (+)- + + - - + + - + - (- )

~------------------------_____________________ + - + - + (+)S-62 - + - + shy(- ) shyS-901 ____________________ - + + - + + + - (- ) (-) shy8-1757 ___________________ - + + - + + + - (- ) - Ii8-1758 ___________________ - + + - + + - + - c-)

S-1759 ___________________ - + + (- ) - + + - + - c-) 8-1760 ___________________ - + + - - + + - + - c-) 8-1761 ___________________ - + C+) - - + + - + - c+) 8-1762 ___________________ - + C+) - - (+) + - + - (+) 8-1763 ___________ - _______ - + + - - + + - + - c-) S-1764 ___________________ - + + - + + - + - (+)

- + + - + + - + - c+)C==)18-1765_ - - bull -___________________ - - - - - - __ - - ---OJ + - + + - (+)8-1766 - + + shy8-1767 ___________________ - + + - + + - + - C+)

- + - - (+) + - (+) - (- )I+ I shy


------------------





Tube 1 Tube 2










cent 22 22 18 22 23 19 20

cent 50 72 34 39 36 30 57

cent 66 81 51 61 62 49 64

cent 73 82 66 73 76 67 70

cent 75 83 76 76 75 77 73

cent 76 88 81 80 76 84 77

cent 0 85 82 81 81 87 80


20 ca 95 96 96 96 96 96


Antibiotic Activi~y







medium 2










Designation used

01 01 1




I I i1





- -







N - ~~ - ~~Czc ~z CQ UJ



l







ci 00 ~



~l il ~l ~l lt~ sectl~~ ~~ g~ ~ ~~ ~~ ~ ~~ 6~~~

~~






I 0




ci 00 ~ g 5 g Cj r- ~~Ogt 0 ~r-




0 0 0

21 0 0

32 0 0

15 1lt 0

0 0 0

0 0 0


0 0 0

18 0 0

26 0 0

(3) 0 0

0 0 0

0 0 0


0 0 0

20 0 0

32 0 0

16 14 0

0 0 0

0 0 0


0 0 0

22 0 0

35 0 0

19 0 0

0 0 0

0 0



0 0

18 18

0 0

0 0

0 0

0 0


0 0

15 14

0 0

(3) (3)

0 0

0 0





-- -- ---- ----











---- 5 7 7 [) 3 0 0 3 4 0









Oritil





Orilin







Orilil





bull

WSI ISW 10 NHCI







































































bull


f

tmiddot

________________

________________



Designation used

NIHT SN-2 (2) __

NIHJ SN-14-L_

NIHJ SN-14-2__

Lilly L _________


ARI1780_______

NI9003 ________

Carpenter_______

E______________

Specific epithet




sp _____________

sp _____________ sp _____________ sp _____________ sp _____________

streptomycini____

vinaceus________


activity

streptomycin______

streptomycin______

streptomycin______

streptomycin_____ _


complex

(1)

(1)

(1)

(1)






Do Do Do







----------------

----------------




activity







---------------




--------------shy




---------------shy

8-1765 _________ ________________ (1) Do8-1766 _________________________ (I) Do8-1767 _________________________ (1) Do






Diastotic Activity








Color






























RESULTS








Diastatic Activity



Nitrate Reduction






Designation used


S-62____________________________ _~~~e~~~~=======================8-901___________________________ _ S-1757__________________________ _ 8-1758__________________________ _8-1759__________________________ _ 8-1760__________________________ _ 8-1761__________________________ _8-1762__________________________ _ 8-1763__________________________ _ 8-1764__________________________ _ 8-1765 __________________________ _ 8-1766__________________________ _8-1767__________________________ _


Tube 1 Tube 2

+ + C+) (+)(+) (+) C+) C+)

(-)

C+) (-)C+) (-)

(+) (+)

(-) (-) (-) (-)

C+) (+)

(+) (+)

(-) (-) (+) (+)

(-)

(-) (-)

(-) (-)

(+) (+)(+) C+)(- ) (- )

C+) (- )(+) (-) (- ) C+) C+)(-) (- )

C+) (+) (- ) (- )

(- )

c+) (+) + +

Organic-based broth

Tube 1 Tube 2

(+) + +

(+) (+) + + - -

(+) (+)c+) (+)

(+) + +

c+) C+)(- ) c+) (+)c-) C-)c+) C+)(- ) c-)(- )

+ +(+) (- ) + + - (+)

(+) + + + + -

(- ) (-) (- )

(- ) (-) (- ) C+) C+)(+) + -C+) C+)

c+) (+) + + + +

C+) c+)c+) c+) + +

(+) c+)

c+) C+)(- ) c-) + + + +

(+) (+)C+) (+) + + + +



- -


B

(p S D


















Tresner name and

color wheel

Pridhllm color

NBS color

NBS color


desigshynation 4

series (28)

series (19)

names of Ridgway

tabs 5

names of CUM tabs 5

















_____

________













































Red 262












01 gt l Il0 a 01 01 0






(- ) + + - + - c+)


Carpenter _________________ - + + (- ) - + + - + - (+)- + + - - + + - + - (- )

~------------------------_____________________ + - + - + (+)S-62 - + - + shy(- ) shyS-901 ____________________ - + + - + + + - (- ) (-) shy8-1757 ___________________ - + + - + + + - (- ) - Ii8-1758 ___________________ - + + - + + - + - c-)

S-1759 ___________________ - + + (- ) - + + - + - c-) 8-1760 ___________________ - + + - - + + - + - c-) 8-1761 ___________________ - + C+) - - + + - + - c+) 8-1762 ___________________ - + C+) - - (+) + - + - (+) 8-1763 ___________ - _______ - + + - - + + - + - c-) S-1764 ___________________ - + + - + + - + - (+)

- + + - + + - + - c+)C==)18-1765_ - - bull -___________________ - - - - - - __ - - ---OJ + - + + - (+)8-1766 - + + shy8-1767 ___________________ - + + - + + - + - C+)

- + - - (+) + - (+) - (- )I+ I shy


------------------





Tube 1 Tube 2










cent 22 22 18 22 23 19 20

cent 50 72 34 39 36 30 57

cent 66 81 51 61 62 49 64

cent 73 82 66 73 76 67 70

cent 75 83 76 76 75 77 73

cent 76 88 81 80 76 84 77

cent 0 85 82 81 81 87 80


20 ca 95 96 96 96 96 96


Antibiotic Activi~y







medium 2










Designation used

01 01 1




I I i1





- -







N - ~~ - ~~Czc ~z CQ UJ



l







ci 00 ~



~l il ~l ~l lt~ sectl~~ ~~ g~ ~ ~~ ~~ ~ ~~ 6~~~

~~






I 0




ci 00 ~ g 5 g Cj r- ~~Ogt 0 ~r-




0 0 0

21 0 0

32 0 0

15 1lt 0

0 0 0

0 0 0


0 0 0

18 0 0

26 0 0

(3) 0 0

0 0 0

0 0 0


0 0 0

20 0 0

32 0 0

16 14 0

0 0 0

0 0 0


0 0 0

22 0 0

35 0 0

19 0 0

0 0 0

0 0



0 0

18 18

0 0

0 0

0 0

0 0


0 0

15 14

0 0

(3) (3)

0 0

0 0





-- -- ---- ----











---- 5 7 7 [) 3 0 0 3 4 0









Oritil





Orilin







Orilil





bull

WSI ISW 10 NHCI







































































bull


f

tmiddot

----------------

----------------




activity







---------------




--------------shy




---------------shy

8-1765 _________ ________________ (1) Do8-1766 _________________________ (I) Do8-1767 _________________________ (1) Do






Diastotic Activity








Color






























RESULTS








Diastatic Activity



Nitrate Reduction






Designation used


S-62____________________________ _~~~e~~~~=======================8-901___________________________ _ S-1757__________________________ _ 8-1758__________________________ _8-1759__________________________ _ 8-1760__________________________ _ 8-1761__________________________ _8-1762__________________________ _ 8-1763__________________________ _ 8-1764__________________________ _ 8-1765 __________________________ _ 8-1766__________________________ _8-1767__________________________ _


Tube 1 Tube 2

+ + C+) (+)(+) (+) C+) C+)

(-)

C+) (-)C+) (-)

(+) (+)

(-) (-) (-) (-)

C+) (+)

(+) (+)

(-) (-) (+) (+)

(-)

(-) (-)

(-) (-)

(+) (+)(+) C+)(- ) (- )

C+) (- )(+) (-) (- ) C+) C+)(-) (- )

C+) (+) (- ) (- )

(- )

c+) (+) + +

Organic-based broth

Tube 1 Tube 2

(+) + +

(+) (+) + + - -

(+) (+)c+) (+)

(+) + +

c+) C+)(- ) c+) (+)c-) C-)c+) C+)(- ) c-)(- )

+ +(+) (- ) + + - (+)

(+) + + + + -

(- ) (-) (- )

(- ) (-) (- ) C+) C+)(+) + -C+) C+)

c+) (+) + + + +

C+) c+)c+) c+) + +

(+) c+)

c+) C+)(- ) c-) + + + +

(+) (+)C+) (+) + + + +



- -


B

(p S D


















Tresner name and

color wheel

Pridhllm color

NBS color

NBS color


desigshynation 4

series (28)

series (19)

names of Ridgway

tabs 5

names of CUM tabs 5

















_____

________













































Red 262












01 gt l Il0 a 01 01 0






(- ) + + - + - c+)


Carpenter _________________ - + + (- ) - + + - + - (+)- + + - - + + - + - (- )

~------------------------_____________________ + - + - + (+)S-62 - + - + shy(- ) shyS-901 ____________________ - + + - + + + - (- ) (-) shy8-1757 ___________________ - + + - + + + - (- ) - Ii8-1758 ___________________ - + + - + + - + - c-)

S-1759 ___________________ - + + (- ) - + + - + - c-) 8-1760 ___________________ - + + - - + + - + - c-) 8-1761 ___________________ - + C+) - - + + - + - c+) 8-1762 ___________________ - + C+) - - (+) + - + - (+) 8-1763 ___________ - _______ - + + - - + + - + - c-) S-1764 ___________________ - + + - + + - + - (+)

- + + - + + - + - c+)C==)18-1765_ - - bull -___________________ - - - - - - __ - - ---OJ + - + + - (+)8-1766 - + + shy8-1767 ___________________ - + + - + + - + - C+)

- + - - (+) + - (+) - (- )I+ I shy


------------------





Tube 1 Tube 2










cent 22 22 18 22 23 19 20

cent 50 72 34 39 36 30 57

cent 66 81 51 61 62 49 64

cent 73 82 66 73 76 67 70

cent 75 83 76 76 75 77 73

cent 76 88 81 80 76 84 77

cent 0 85 82 81 81 87 80


20 ca 95 96 96 96 96 96


Antibiotic Activi~y







medium 2










Designation used

01 01 1




I I i1





- -







N - ~~ - ~~Czc ~z CQ UJ



l







ci 00 ~



~l il ~l ~l lt~ sectl~~ ~~ g~ ~ ~~ ~~ ~ ~~ 6~~~

~~






I 0




ci 00 ~ g 5 g Cj r- ~~Ogt 0 ~r-




0 0 0

21 0 0

32 0 0

15 1lt 0

0 0 0

0 0 0


0 0 0

18 0 0

26 0 0

(3) 0 0

0 0 0

0 0 0


0 0 0

20 0 0

32 0 0

16 14 0

0 0 0

0 0 0


0 0 0

22 0 0

35 0 0

19 0 0

0 0 0

0 0



0 0

18 18

0 0

0 0

0 0

0 0


0 0

15 14

0 0

(3) (3)

0 0

0 0





-- -- ---- ----











---- 5 7 7 [) 3 0 0 3 4 0









Oritil





Orilin







Orilil





bull

WSI ISW 10 NHCI







































































bull


f

tmiddot



Diastotic Activity








Color






























RESULTS








Diastatic Activity



Nitrate Reduction






Designation used


S-62____________________________ _~~~e~~~~=======================8-901___________________________ _ S-1757__________________________ _ 8-1758__________________________ _8-1759__________________________ _ 8-1760__________________________ _ 8-1761__________________________ _8-1762__________________________ _ 8-1763__________________________ _ 8-1764__________________________ _ 8-1765 __________________________ _ 8-1766__________________________ _8-1767__________________________ _


Tube 1 Tube 2

+ + C+) (+)(+) (+) C+) C+)

(-)

C+) (-)C+) (-)

(+) (+)

(-) (-) (-) (-)

C+) (+)

(+) (+)

(-) (-) (+) (+)

(-)

(-) (-)

(-) (-)

(+) (+)(+) C+)(- ) (- )

C+) (- )(+) (-) (- ) C+) C+)(-) (- )

C+) (+) (- ) (- )

(- )

c+) (+) + +

Organic-based broth

Tube 1 Tube 2

(+) + +

(+) (+) + + - -

(+) (+)c+) (+)

(+) + +

c+) C+)(- ) c+) (+)c-) C-)c+) C+)(- ) c-)(- )

+ +(+) (- ) + + - (+)

(+) + + + + -

(- ) (-) (- )

(- ) (-) (- ) C+) C+)(+) + -C+) C+)

c+) (+) + + + +

C+) c+)c+) c+) + +

(+) c+)

c+) C+)(- ) c-) + + + +

(+) (+)C+) (+) + + + +



- -


B

(p S D


















Tresner name and

color wheel

Pridhllm color

NBS color

NBS color


desigshynation 4

series (28)

series (19)

names of Ridgway

tabs 5

names of CUM tabs 5

















_____

________













































Red 262












01 gt l Il0 a 01 01 0






(- ) + + - + - c+)


Carpenter _________________ - + + (- ) - + + - + - (+)- + + - - + + - + - (- )

~------------------------_____________________ + - + - + (+)S-62 - + - + shy(- ) shyS-901 ____________________ - + + - + + + - (- ) (-) shy8-1757 ___________________ - + + - + + + - (- ) - Ii8-1758 ___________________ - + + - + + - + - c-)

S-1759 ___________________ - + + (- ) - + + - + - c-) 8-1760 ___________________ - + + - - + + - + - c-) 8-1761 ___________________ - + C+) - - + + - + - c+) 8-1762 ___________________ - + C+) - - (+) + - + - (+) 8-1763 ___________ - _______ - + + - - + + - + - c-) S-1764 ___________________ - + + - + + - + - (+)

- + + - + + - + - c+)C==)18-1765_ - - bull -___________________ - - - - - - __ - - ---OJ + - + + - (+)8-1766 - + + shy8-1767 ___________________ - + + - + + - + - C+)

- + - - (+) + - (+) - (- )I+ I shy


------------------





Tube 1 Tube 2










cent 22 22 18 22 23 19 20

cent 50 72 34 39 36 30 57

cent 66 81 51 61 62 49 64

cent 73 82 66 73 76 67 70

cent 75 83 76 76 75 77 73

cent 76 88 81 80 76 84 77

cent 0 85 82 81 81 87 80


20 ca 95 96 96 96 96 96


Antibiotic Activi~y







medium 2










Designation used

01 01 1




I I i1





- -







N - ~~ - ~~Czc ~z CQ UJ



l







ci 00 ~



~l il ~l ~l lt~ sectl~~ ~~ g~ ~ ~~ ~~ ~ ~~ 6~~~

~~






I 0




ci 00 ~ g 5 g Cj r- ~~Ogt 0 ~r-




0 0 0

21 0 0

32 0 0

15 1lt 0

0 0 0

0 0 0


0 0 0

18 0 0

26 0 0

(3) 0 0

0 0 0

0 0 0


0 0 0

20 0 0

32 0 0

16 14 0

0 0 0

0 0 0


0 0 0

22 0 0

35 0 0

19 0 0

0 0 0

0 0



0 0

18 18

0 0

0 0

0 0

0 0


0 0

15 14

0 0

(3) (3)

0 0

0 0





-- -- ---- ----











---- 5 7 7 [) 3 0 0 3 4 0









Oritil





Orilin







Orilil





bull

WSI ISW 10 NHCI







































































bull


f

tmiddot



























RESULTS








Diastatic Activity



Nitrate Reduction






Designation used


S-62____________________________ _~~~e~~~~=======================8-901___________________________ _ S-1757__________________________ _ 8-1758__________________________ _8-1759__________________________ _ 8-1760__________________________ _ 8-1761__________________________ _8-1762__________________________ _ 8-1763__________________________ _ 8-1764__________________________ _ 8-1765 __________________________ _ 8-1766__________________________ _8-1767__________________________ _


Tube 1 Tube 2

+ + C+) (+)(+) (+) C+) C+)

(-)

C+) (-)C+) (-)

(+) (+)

(-) (-) (-) (-)

C+) (+)

(+) (+)

(-) (-) (+) (+)

(-)

(-) (-)

(-) (-)

(+) (+)(+) C+)(- ) (- )

C+) (- )(+) (-) (- ) C+) C+)(-) (- )

C+) (+) (- ) (- )

(- )

c+) (+) + +

Organic-based broth

Tube 1 Tube 2

(+) + +

(+) (+) + + - -

(+) (+)c+) (+)

(+) + +

c+) C+)(- ) c+) (+)c-) C-)c+) C+)(- ) c-)(- )

+ +(+) (- ) + + - (+)

(+) + + + + -

(- ) (-) (- )

(- ) (-) (- ) C+) C+)(+) + -C+) C+)

c+) (+) + + + +

C+) c+)c+) c+) + +

(+) c+)

c+) C+)(- ) c-) + + + +

(+) (+)C+) (+) + + + +



- -


B

(p S D


















Tresner name and

color wheel

Pridhllm color

NBS color

NBS color


desigshynation 4

series (28)

series (19)

names of Ridgway

tabs 5

names of CUM tabs 5

















_____

________













































Red 262












01 gt l Il0 a 01 01 0






(- ) + + - + - c+)


Carpenter _________________ - + + (- ) - + + - + - (+)- + + - - + + - + - (- )

~------------------------_____________________ + - + - + (+)S-62 - + - + shy(- ) shyS-901 ____________________ - + + - + + + - (- ) (-) shy8-1757 ___________________ - + + - + + + - (- ) - Ii8-1758 ___________________ - + + - + + - + - c-)

S-1759 ___________________ - + + (- ) - + + - + - c-) 8-1760 ___________________ - + + - - + + - + - c-) 8-1761 ___________________ - + C+) - - + + - + - c+) 8-1762 ___________________ - + C+) - - (+) + - + - (+) 8-1763 ___________ - _______ - + + - - + + - + - c-) S-1764 ___________________ - + + - + + - + - (+)

- + + - + + - + - c+)C==)18-1765_ - - bull -___________________ - - - - - - __ - - ---OJ + - + + - (+)8-1766 - + + shy8-1767 ___________________ - + + - + + - + - C+)

- + - - (+) + - (+) - (- )I+ I shy


------------------





Tube 1 Tube 2










cent 22 22 18 22 23 19 20

cent 50 72 34 39 36 30 57

cent 66 81 51 61 62 49 64

cent 73 82 66 73 76 67 70

cent 75 83 76 76 75 77 73

cent 76 88 81 80 76 84 77

cent 0 85 82 81 81 87 80


20 ca 95 96 96 96 96 96


Antibiotic Activi~y







medium 2










Designation used

01 01 1




I I i1





- -







N - ~~ - ~~Czc ~z CQ UJ



l







ci 00 ~



~l il ~l ~l lt~ sectl~~ ~~ g~ ~ ~~ ~~ ~ ~~ 6~~~

~~






I 0




ci 00 ~ g 5 g Cj r- ~~Ogt 0 ~r-




0 0 0

21 0 0

32 0 0

15 1lt 0

0 0 0

0 0 0


0 0 0

18 0 0

26 0 0

(3) 0 0

0 0 0

0 0 0


0 0 0

20 0 0

32 0 0

16 14 0

0 0 0

0 0 0


0 0 0

22 0 0

35 0 0

19 0 0

0 0 0

0 0



0 0

18 18

0 0

0 0

0 0

0 0


0 0

15 14

0 0

(3) (3)

0 0

0 0





-- -- ---- ----











---- 5 7 7 [) 3 0 0 3 4 0









Oritil





Orilin







Orilil





bull

WSI ISW 10 NHCI







































































bull


f

tmiddot




Diastatic Activity



Nitrate Reduction






Designation used


S-62____________________________ _~~~e~~~~=======================8-901___________________________ _ S-1757__________________________ _ 8-1758__________________________ _8-1759__________________________ _ 8-1760__________________________ _ 8-1761__________________________ _8-1762__________________________ _ 8-1763__________________________ _ 8-1764__________________________ _ 8-1765 __________________________ _ 8-1766__________________________ _8-1767__________________________ _


Tube 1 Tube 2

+ + C+) (+)(+) (+) C+) C+)

(-)

C+) (-)C+) (-)

(+) (+)

(-) (-) (-) (-)

C+) (+)

(+) (+)

(-) (-) (+) (+)

(-)

(-) (-)

(-) (-)

(+) (+)(+) C+)(- ) (- )

C+) (- )(+) (-) (- ) C+) C+)(-) (- )

C+) (+) (- ) (- )

(- )

c+) (+) + +

Organic-based broth

Tube 1 Tube 2

(+) + +

(+) (+) + + - -

(+) (+)c+) (+)

(+) + +

c+) C+)(- ) c+) (+)c-) C-)c+) C+)(- ) c-)(- )

+ +(+) (- ) + + - (+)

(+) + + + + -

(- ) (-) (- )

(- ) (-) (- ) C+) C+)(+) + -C+) C+)

c+) (+) + + + +

C+) c+)c+) c+) + +

(+) c+)

c+) C+)(- ) c-) + + + +

(+) (+)C+) (+) + + + +



- -


B

(p S D


















Tresner name and

color wheel

Pridhllm color

NBS color

NBS color


desigshynation 4

series (28)

series (19)

names of Ridgway

tabs 5

names of CUM tabs 5

















_____

________













































Red 262












01 gt l Il0 a 01 01 0






(- ) + + - + - c+)


Carpenter _________________ - + + (- ) - + + - + - (+)- + + - - + + - + - (- )

~------------------------_____________________ + - + - + (+)S-62 - + - + shy(- ) shyS-901 ____________________ - + + - + + + - (- ) (-) shy8-1757 ___________________ - + + - + + + - (- ) - Ii8-1758 ___________________ - + + - + + - + - c-)

S-1759 ___________________ - + + (- ) - + + - + - c-) 8-1760 ___________________ - + + - - + + - + - c-) 8-1761 ___________________ - + C+) - - + + - + - c+) 8-1762 ___________________ - + C+) - - (+) + - + - (+) 8-1763 ___________ - _______ - + + - - + + - + - c-) S-1764 ___________________ - + + - + + - + - (+)

- + + - + + - + - c+)C==)18-1765_ - - bull -___________________ - - - - - - __ - - ---OJ + - + + - (+)8-1766 - + + shy8-1767 ___________________ - + + - + + - + - C+)

- + - - (+) + - (+) - (- )I+ I shy


------------------





Tube 1 Tube 2










cent 22 22 18 22 23 19 20

cent 50 72 34 39 36 30 57

cent 66 81 51 61 62 49 64

cent 73 82 66 73 76 67 70

cent 75 83 76 76 75 77 73

cent 76 88 81 80 76 84 77

cent 0 85 82 81 81 87 80


20 ca 95 96 96 96 96 96


Antibiotic Activi~y







medium 2










Designation used

01 01 1




I I i1





- -







N - ~~ - ~~Czc ~z CQ UJ



l







ci 00 ~



~l il ~l ~l lt~ sectl~~ ~~ g~ ~ ~~ ~~ ~ ~~ 6~~~

~~






I 0




ci 00 ~ g 5 g Cj r- ~~Ogt 0 ~r-




0 0 0

21 0 0

32 0 0

15 1lt 0

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(3) 0 0

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0 0

18 18

0 0

0 0

0 0

0 0


0 0

15 14

0 0

(3) (3)

0 0

0 0





-- -- ---- ----











---- 5 7 7 [) 3 0 0 3 4 0









Oritil





Orilin







Orilil





bull

WSI ISW 10 NHCI







































































bull


f

tmiddot



Designation used


S-62____________________________ _~~~e~~~~=======================8-901___________________________ _ S-1757__________________________ _ 8-1758__________________________ _8-1759__________________________ _ 8-1760__________________________ _ 8-1761__________________________ _8-1762__________________________ _ 8-1763__________________________ _ 8-1764__________________________ _ 8-1765 __________________________ _ 8-1766__________________________ _8-1767__________________________ _


Tube 1 Tube 2

+ + C+) (+)(+) (+) C+) C+)

(-)

C+) (-)C+) (-)

(+) (+)

(-) (-) (-) (-)

C+) (+)

(+) (+)

(-) (-) (+) (+)

(-)

(-) (-)

(-) (-)

(+) (+)(+) C+)(- ) (- )

C+) (- )(+) (-) (- ) C+) C+)(-) (- )

C+) (+) (- ) (- )

(- )

c+) (+) + +

Organic-based broth

Tube 1 Tube 2

(+) + +

(+) (+) + + - -

(+) (+)c+) (+)

(+) + +

c+) C+)(- ) c+) (+)c-) C-)c+) C+)(- ) c-)(- )

+ +(+) (- ) + + - (+)

(+) + + + + -

(- ) (-) (- )

(- ) (-) (- ) C+) C+)(+) + -C+) C+)

c+) (+) + + + +

C+) c+)c+) c+) + +

(+) c+)

c+) C+)(- ) c-) + + + +

(+) (+)C+) (+) + + + +



- -


B

(p S D


















Tresner name and

color wheel

Pridhllm color

NBS color

NBS color


desigshynation 4

series (28)

series (19)

names of Ridgway

tabs 5

names of CUM tabs 5

















_____

________













































Red 262












01 gt l Il0 a 01 01 0






(- ) + + - + - c+)


Carpenter _________________ - + + (- ) - + + - + - (+)- + + - - + + - + - (- )

~------------------------_____________________ + - + - + (+)S-62 - + - + shy(- ) shyS-901 ____________________ - + + - + + + - (- ) (-) shy8-1757 ___________________ - + + - + + + - (- ) - Ii8-1758 ___________________ - + + - + + - + - c-)

S-1759 ___________________ - + + (- ) - + + - + - c-) 8-1760 ___________________ - + + - - + + - + - c-) 8-1761 ___________________ - + C+) - - + + - + - c+) 8-1762 ___________________ - + C+) - - (+) + - + - (+) 8-1763 ___________ - _______ - + + - - + + - + - c-) S-1764 ___________________ - + + - + + - + - (+)

- + + - + + - + - c+)C==)18-1765_ - - bull -___________________ - - - - - - __ - - ---OJ + - + + - (+)8-1766 - + + shy8-1767 ___________________ - + + - + + - + - C+)

- + - - (+) + - (+) - (- )I+ I shy


------------------





Tube 1 Tube 2










cent 22 22 18 22 23 19 20

cent 50 72 34 39 36 30 57

cent 66 81 51 61 62 49 64

cent 73 82 66 73 76 67 70

cent 75 83 76 76 75 77 73

cent 76 88 81 80 76 84 77

cent 0 85 82 81 81 87 80


20 ca 95 96 96 96 96 96


Antibiotic Activi~y







medium 2










Designation used

01 01 1




I I i1





- -







N - ~~ - ~~Czc ~z CQ UJ



l







ci 00 ~



~l il ~l ~l lt~ sectl~~ ~~ g~ ~ ~~ ~~ ~ ~~ 6~~~

~~






I 0




ci 00 ~ g 5 g Cj r- ~~Ogt 0 ~r-




0 0 0

21 0 0

32 0 0

15 1lt 0

0 0 0

0 0 0


0 0 0

18 0 0

26 0 0

(3) 0 0

0 0 0

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16 14 0

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0 0 0

22 0 0

35 0 0

19 0 0

0 0 0

0 0



0 0

18 18

0 0

0 0

0 0

0 0


0 0

15 14

0 0

(3) (3)

0 0

0 0





-- -- ---- ----











---- 5 7 7 [) 3 0 0 3 4 0









Oritil





Orilin







Orilil





bull

WSI ISW 10 NHCI







































































bull


f

tmiddot


- -


B

(p S D


















Tresner name and

color wheel

Pridhllm color

NBS color

NBS color


desigshynation 4

series (28)

series (19)

names of Ridgway

tabs 5

names of CUM tabs 5

















_____

________













































Red 262












01 gt l Il0 a 01 01 0






(- ) + + - + - c+)


Carpenter _________________ - + + (- ) - + + - + - (+)- + + - - + + - + - (- )

~------------------------_____________________ + - + - + (+)S-62 - + - + shy(- ) shyS-901 ____________________ - + + - + + + - (- ) (-) shy8-1757 ___________________ - + + - + + + - (- ) - Ii8-1758 ___________________ - + + - + + - + - c-)

S-1759 ___________________ - + + (- ) - + + - + - c-) 8-1760 ___________________ - + + - - + + - + - c-) 8-1761 ___________________ - + C+) - - + + - + - c+) 8-1762 ___________________ - + C+) - - (+) + - + - (+) 8-1763 ___________ - _______ - + + - - + + - + - c-) S-1764 ___________________ - + + - + + - + - (+)

- + + - + + - + - c+)C==)18-1765_ - - bull -___________________ - - - - - - __ - - ---OJ + - + + - (+)8-1766 - + + shy8-1767 ___________________ - + + - + + - + - C+)

- + - - (+) + - (+) - (- )I+ I shy


------------------





Tube 1 Tube 2










cent 22 22 18 22 23 19 20

cent 50 72 34 39 36 30 57

cent 66 81 51 61 62 49 64

cent 73 82 66 73 76 67 70

cent 75 83 76 76 75 77 73

cent 76 88 81 80 76 84 77

cent 0 85 82 81 81 87 80


20 ca 95 96 96 96 96 96


Antibiotic Activi~y







medium 2










Designation used

01 01 1




I I i1





- -







N - ~~ - ~~Czc ~z CQ UJ



l







ci 00 ~



~l il ~l ~l lt~ sectl~~ ~~ g~ ~ ~~ ~~ ~ ~~ 6~~~

~~






I 0




ci 00 ~ g 5 g Cj r- ~~Ogt 0 ~r-




0 0 0

21 0 0

32 0 0

15 1lt 0

0 0 0

0 0 0


0 0 0

18 0 0

26 0 0

(3) 0 0

0 0 0

0 0 0


0 0 0

20 0 0

32 0 0

16 14 0

0 0 0

0 0 0


0 0 0

22 0 0

35 0 0

19 0 0

0 0 0

0 0



0 0

18 18

0 0

0 0

0 0

0 0


0 0

15 14

0 0

(3) (3)

0 0

0 0





-- -- ---- ----











---- 5 7 7 [) 3 0 0 3 4 0









Oritil





Orilin







Orilil





bull

WSI ISW 10 NHCI







































































bull


f

tmiddot











Tresner name and

color wheel

Pridhllm color

NBS color

NBS color


desigshynation 4

series (28)

series (19)

names of Ridgway

tabs 5

names of CUM tabs 5

















_____

________













































Red 262












01 gt l Il0 a 01 01 0






(- ) + + - + - c+)


Carpenter _________________ - + + (- ) - + + - + - (+)- + + - - + + - + - (- )

~------------------------_____________________ + - + - + (+)S-62 - + - + shy(- ) shyS-901 ____________________ - + + - + + + - (- ) (-) shy8-1757 ___________________ - + + - + + + - (- ) - Ii8-1758 ___________________ - + + - + + - + - c-)

S-1759 ___________________ - + + (- ) - + + - + - c-) 8-1760 ___________________ - + + - - + + - + - c-) 8-1761 ___________________ - + C+) - - + + - + - c+) 8-1762 ___________________ - + C+) - - (+) + - + - (+) 8-1763 ___________ - _______ - + + - - + + - + - c-) S-1764 ___________________ - + + - + + - + - (+)

- + + - + + - + - c+)C==)18-1765_ - - bull -___________________ - - - - - - __ - - ---OJ + - + + - (+)8-1766 - + + shy8-1767 ___________________ - + + - + + - + - C+)

- + - - (+) + - (+) - (- )I+ I shy


------------------





Tube 1 Tube 2










cent 22 22 18 22 23 19 20

cent 50 72 34 39 36 30 57

cent 66 81 51 61 62 49 64

cent 73 82 66 73 76 67 70

cent 75 83 76 76 75 77 73

cent 76 88 81 80 76 84 77

cent 0 85 82 81 81 87 80


20 ca 95 96 96 96 96 96


Antibiotic Activi~y







medium 2










Designation used

01 01 1




I I i1





- -







N - ~~ - ~~Czc ~z CQ UJ



l







ci 00 ~



~l il ~l ~l lt~ sectl~~ ~~ g~ ~ ~~ ~~ ~ ~~ 6~~~

~~






I 0




ci 00 ~ g 5 g Cj r- ~~Ogt 0 ~r-




0 0 0

21 0 0

32 0 0

15 1lt 0

0 0 0

0 0 0


0 0 0

18 0 0

26 0 0

(3) 0 0

0 0 0

0 0 0


0 0 0

20 0 0

32 0 0

16 14 0

0 0 0

0 0 0


0 0 0

22 0 0

35 0 0

19 0 0

0 0 0

0 0



0 0

18 18

0 0

0 0

0 0

0 0


0 0

15 14

0 0

(3) (3)

0 0

0 0





-- -- ---- ----











---- 5 7 7 [) 3 0 0 3 4 0









Oritil





Orilin







Orilil





bull

WSI ISW 10 NHCI







































































bull


f

tmiddot

_____

________













































Red 262












01 gt l Il0 a 01 01 0






(- ) + + - + - c+)


Carpenter _________________ - + + (- ) - + + - + - (+)- + + - - + + - + - (- )

~------------------------_____________________ + - + - + (+)S-62 - + - + shy(- ) shyS-901 ____________________ - + + - + + + - (- ) (-) shy8-1757 ___________________ - + + - + + + - (- ) - Ii8-1758 ___________________ - + + - + + - + - c-)

S-1759 ___________________ - + + (- ) - + + - + - c-) 8-1760 ___________________ - + + - - + + - + - c-) 8-1761 ___________________ - + C+) - - + + - + - c+) 8-1762 ___________________ - + C+) - - (+) + - + - (+) 8-1763 ___________ - _______ - + + - - + + - + - c-) S-1764 ___________________ - + + - + + - + - (+)

- + + - + + - + - c+)C==)18-1765_ - - bull -___________________ - - - - - - __ - - ---OJ + - + + - (+)8-1766 - + + shy8-1767 ___________________ - + + - + + - + - C+)

- + - - (+) + - (+) - (- )I+ I shy


------------------





Tube 1 Tube 2










cent 22 22 18 22 23 19 20

cent 50 72 34 39 36 30 57

cent 66 81 51 61 62 49 64

cent 73 82 66 73 76 67 70

cent 75 83 76 76 75 77 73

cent 76 88 81 80 76 84 77

cent 0 85 82 81 81 87 80


20 ca 95 96 96 96 96 96


Antibiotic Activi~y







medium 2










Designation used

01 01 1




I I i1





- -







N - ~~ - ~~Czc ~z CQ UJ



l







ci 00 ~



~l il ~l ~l lt~ sectl~~ ~~ g~ ~ ~~ ~~ ~ ~~ 6~~~

~~






I 0




ci 00 ~ g 5 g Cj r- ~~Ogt 0 ~r-




0 0 0

21 0 0

32 0 0

15 1lt 0

0 0 0

0 0 0


0 0 0

18 0 0

26 0 0

(3) 0 0

0 0 0

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20 0 0

32 0 0

16 14 0

0 0 0

0 0 0


0 0 0

22 0 0

35 0 0

19 0 0

0 0 0

0 0



0 0

18 18

0 0

0 0

0 0

0 0


0 0

15 14

0 0

(3) (3)

0 0

0 0





-- -- ---- ----











---- 5 7 7 [) 3 0 0 3 4 0









Oritil





Orilin







Orilil





bull

WSI ISW 10 NHCI







































































bull


f

tmiddot



















Red 262












01 gt l Il0 a 01 01 0






(- ) + + - + - c+)


Carpenter _________________ - + + (- ) - + + - + - (+)- + + - - + + - + - (- )

~------------------------_____________________ + - + - + (+)S-62 - + - + shy(- ) shyS-901 ____________________ - + + - + + + - (- ) (-) shy8-1757 ___________________ - + + - + + + - (- ) - Ii8-1758 ___________________ - + + - + + - + - c-)

S-1759 ___________________ - + + (- ) - + + - + - c-) 8-1760 ___________________ - + + - - + + - + - c-) 8-1761 ___________________ - + C+) - - + + - + - c+) 8-1762 ___________________ - + C+) - - (+) + - + - (+) 8-1763 ___________ - _______ - + + - - + + - + - c-) S-1764 ___________________ - + + - + + - + - (+)

- + + - + + - + - c+)C==)18-1765_ - - bull -___________________ - - - - - - __ - - ---OJ + - + + - (+)8-1766 - + + shy8-1767 ___________________ - + + - + + - + - C+)

- + - - (+) + - (+) - (- )I+ I shy


------------------





Tube 1 Tube 2










cent 22 22 18 22 23 19 20

cent 50 72 34 39 36 30 57

cent 66 81 51 61 62 49 64

cent 73 82 66 73 76 67 70

cent 75 83 76 76 75 77 73

cent 76 88 81 80 76 84 77

cent 0 85 82 81 81 87 80


20 ca 95 96 96 96 96 96


Antibiotic Activi~y







medium 2










Designation used

01 01 1




I I i1





- -







N - ~~ - ~~Czc ~z CQ UJ



l







ci 00 ~



~l il ~l ~l lt~ sectl~~ ~~ g~ ~ ~~ ~~ ~ ~~ 6~~~

~~






I 0




ci 00 ~ g 5 g Cj r- ~~Ogt 0 ~r-




0 0 0

21 0 0

32 0 0

15 1lt 0

0 0 0

0 0 0


0 0 0

18 0 0

26 0 0

(3) 0 0

0 0 0

0 0 0


0 0 0

20 0 0

32 0 0

16 14 0

0 0 0

0 0 0


0 0 0

22 0 0

35 0 0

19 0 0

0 0 0

0 0



0 0

18 18

0 0

0 0

0 0

0 0


0 0

15 14

0 0

(3) (3)

0 0

0 0





-- -- ---- ----











---- 5 7 7 [) 3 0 0 3 4 0









Oritil





Orilin







Orilil





bull

WSI ISW 10 NHCI







































































bull


f

tmiddot




01 gt l Il0 a 01 01 0






(- ) + + - + - c+)


Carpenter _________________ - + + (- ) - + + - + - (+)- + + - - + + - + - (- )

~------------------------_____________________ + - + - + (+)S-62 - + - + shy(- ) shyS-901 ____________________ - + + - + + + - (- ) (-) shy8-1757 ___________________ - + + - + + + - (- ) - Ii8-1758 ___________________ - + + - + + - + - c-)

S-1759 ___________________ - + + (- ) - + + - + - c-) 8-1760 ___________________ - + + - - + + - + - c-) 8-1761 ___________________ - + C+) - - + + - + - c+) 8-1762 ___________________ - + C+) - - (+) + - + - (+) 8-1763 ___________ - _______ - + + - - + + - + - c-) S-1764 ___________________ - + + - + + - + - (+)

- + + - + + - + - c+)C==)18-1765_ - - bull -___________________ - - - - - - __ - - ---OJ + - + + - (+)8-1766 - + + shy8-1767 ___________________ - + + - + + - + - C+)

- + - - (+) + - (+) - (- )I+ I shy


------------------





Tube 1 Tube 2










cent 22 22 18 22 23 19 20

cent 50 72 34 39 36 30 57

cent 66 81 51 61 62 49 64

cent 73 82 66 73 76 67 70

cent 75 83 76 76 75 77 73

cent 76 88 81 80 76 84 77

cent 0 85 82 81 81 87 80


20 ca 95 96 96 96 96 96


Antibiotic Activi~y







medium 2










Designation used

01 01 1




I I i1





- -







N - ~~ - ~~Czc ~z CQ UJ



l







ci 00 ~



~l il ~l ~l lt~ sectl~~ ~~ g~ ~ ~~ ~~ ~ ~~ 6~~~

~~






I 0




ci 00 ~ g 5 g Cj r- ~~Ogt 0 ~r-




0 0 0

21 0 0

32 0 0

15 1lt 0

0 0 0

0 0 0


0 0 0

18 0 0

26 0 0

(3) 0 0

0 0 0

0 0 0


0 0 0

20 0 0

32 0 0

16 14 0

0 0 0

0 0 0


0 0 0

22 0 0

35 0 0

19 0 0

0 0 0

0 0



0 0

18 18

0 0

0 0

0 0

0 0


0 0

15 14

0 0

(3) (3)

0 0

0 0





-- -- ---- ----











---- 5 7 7 [) 3 0 0 3 4 0









Oritil





Orilin







Orilil





bull

WSI ISW 10 NHCI







































































bull


f

tmiddot

------------------





Tube 1 Tube 2










cent 22 22 18 22 23 19 20

cent 50 72 34 39 36 30 57

cent 66 81 51 61 62 49 64

cent 73 82 66 73 76 67 70

cent 75 83 76 76 75 77 73

cent 76 88 81 80 76 84 77

cent 0 85 82 81 81 87 80


20 ca 95 96 96 96 96 96


Antibiotic Activi~y







medium 2










Designation used

01 01 1




I I i1





- -







N - ~~ - ~~Czc ~z CQ UJ



l







ci 00 ~



~l il ~l ~l lt~ sectl~~ ~~ g~ ~ ~~ ~~ ~ ~~ 6~~~

~~






I 0




ci 00 ~ g 5 g Cj r- ~~Ogt 0 ~r-




0 0 0

21 0 0

32 0 0

15 1lt 0

0 0 0

0 0 0


0 0 0

18 0 0

26 0 0

(3) 0 0

0 0 0

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0 0 0

20 0 0

32 0 0

16 14 0

0 0 0

0 0 0


0 0 0

22 0 0

35 0 0

19 0 0

0 0 0

0 0



0 0

18 18

0 0

0 0

0 0

0 0


0 0

15 14

0 0

(3) (3)

0 0

0 0





-- -- ---- ----











---- 5 7 7 [) 3 0 0 3 4 0









Oritil





Orilin







Orilil





bull

WSI ISW 10 NHCI







































































bull


f

tmiddot




medium 2










Designation used

01 01 1




I I i1





- -







N - ~~ - ~~Czc ~z CQ UJ



l







ci 00 ~



~l il ~l ~l lt~ sectl~~ ~~ g~ ~ ~~ ~~ ~ ~~ 6~~~

~~






I 0




ci 00 ~ g 5 g Cj r- ~~Ogt 0 ~r-




0 0 0

21 0 0

32 0 0

15 1lt 0

0 0 0

0 0 0


0 0 0

18 0 0

26 0 0

(3) 0 0

0 0 0

0 0 0


0 0 0

20 0 0

32 0 0

16 14 0

0 0 0

0 0 0


0 0 0

22 0 0

35 0 0

19 0 0

0 0 0

0 0



0 0

18 18

0 0

0 0

0 0

0 0


0 0

15 14

0 0

(3) (3)

0 0

0 0





-- -- ---- ----











---- 5 7 7 [) 3 0 0 3 4 0









Oritil





Orilin







Orilil





bull

WSI ISW 10 NHCI







































































bull


f

tmiddot



Designation used

01 01 1




I I i1





- -







N - ~~ - ~~Czc ~z CQ UJ



l







ci 00 ~



~l il ~l ~l lt~ sectl~~ ~~ g~ ~ ~~ ~~ ~ ~~ 6~~~

~~






I 0




ci 00 ~ g 5 g Cj r- ~~Ogt 0 ~r-




0 0 0

21 0 0

32 0 0

15 1lt 0

0 0 0

0 0 0


0 0 0

18 0 0

26 0 0

(3) 0 0

0 0 0

0 0 0


0 0 0

20 0 0

32 0 0

16 14 0

0 0 0

0 0 0


0 0 0

22 0 0

35 0 0

19 0 0

0 0 0

0 0



0 0

18 18

0 0

0 0

0 0

0 0


0 0

15 14

0 0

(3) (3)

0 0

0 0





-- -- ---- ----











---- 5 7 7 [) 3 0 0 3 4 0









Oritil





Orilin







Orilil





bull

WSI ISW 10 NHCI







































































bull


f

tmiddot

- -







N - ~~ - ~~Czc ~z CQ UJ



l







ci 00 ~



~l il ~l ~l lt~ sectl~~ ~~ g~ ~ ~~ ~~ ~ ~~ 6~~~

~~






I 0




ci 00 ~ g 5 g Cj r- ~~Ogt 0 ~r-




0 0 0

21 0 0

32 0 0

15 1lt 0

0 0 0

0 0 0


0 0 0

18 0 0

26 0 0

(3) 0 0

0 0 0

0 0 0


0 0 0

20 0 0

32 0 0

16 14 0

0 0 0

0 0 0


0 0 0

22 0 0

35 0 0

19 0 0

0 0 0

0 0



0 0

18 18

0 0

0 0

0 0

0 0


0 0

15 14

0 0

(3) (3)

0 0

0 0





-- -- ---- ----











---- 5 7 7 [) 3 0 0 3 4 0









Oritil





Orilin







Orilil





bull

WSI ISW 10 NHCI







































































bull


f

tmiddot







ci 00 ~



~l il ~l ~l lt~ sectl~~ ~~ g~ ~ ~~ ~~ ~ ~~ 6~~~

~~






I 0




ci 00 ~ g 5 g Cj r- ~~Ogt 0 ~r-




0 0 0

21 0 0

32 0 0

15 1lt 0

0 0 0

0 0 0


0 0 0

18 0 0

26 0 0

(3) 0 0

0 0 0

0 0 0


0 0 0

20 0 0

32 0 0

16 14 0

0 0 0

0 0 0


0 0 0

22 0 0

35 0 0

19 0 0

0 0 0

0 0



0 0

18 18

0 0

0 0

0 0

0 0


0 0

15 14

0 0

(3) (3)

0 0

0 0





-- -- ---- ----











---- 5 7 7 [) 3 0 0 3 4 0









Oritil





Orilin







Orilil





bull

WSI ISW 10 NHCI







































































bull


f

tmiddot



ci 00 ~ g 5 g Cj r- ~~Ogt 0 ~r-




0 0 0

21 0 0

32 0 0

15 1lt 0

0 0 0

0 0 0


0 0 0

18 0 0

26 0 0

(3) 0 0

0 0 0

0 0 0


0 0 0

20 0 0

32 0 0

16 14 0

0 0 0

0 0 0


0 0 0

22 0 0

35 0 0

19 0 0

0 0 0

0 0



0 0

18 18

0 0

0 0

0 0

0 0


0 0

15 14

0 0

(3) (3)

0 0

0 0





-- -- ---- ----











---- 5 7 7 [) 3 0 0 3 4 0









Oritil





Orilin







Orilil





bull

WSI ISW 10 NHCI







































































bull


f

tmiddot

-- -- ---- ----











---- 5 7 7 [) 3 0 0 3 4 0









Oritil





Orilin







Orilil





bull

WSI ISW 10 NHCI







































































bull


f

tmiddot


Oritil





Orilin







Orilil





bull

WSI ISW 10 NHCI







































































bull


f

tmiddot


Orilil





bull

WSI ISW 10 NHCI







































































bull


f

tmiddot




































































bull


f

tmiddot

f

tmiddot

Streptomyces Griseus (Krainsky) Waksman and Henrici: A Taxonomic...

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Transcript of Streptomyces Griseus (Krainsky) Waksman and Henrici: A Taxonomic...